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Buffalo vs Chicago Recap - Blackhawks Win 4-3
Buffalo was unable to break down Marty Turco on Saturday, firing 41 shots but scoring only 3 times in a 4-3 loss against the Blackhawks.
Chicago won as a -178 favorite, while Over-Under bettors who played OVER the total of 5.5 were rewarded at the payout window.
Chicago got goals from Patrick Sharp (2), Marian Hossa, and Dave Bolland in the victory. Sharp opened the scoring at 6:25 of the first period.
Marty Turco stopped 38 of 41 Buffalo shots in the victory.
Buffalo got goals from Cody McCormick, Paul Gaustad, and Drew Stafford in the loss, but it wasn't enough.
Patrick Lalime stopped 17 of 21 shots for the Sabres, but it wasn't enough to earn the victory.
On the power play, Chicago went 1-for-4, while the Sabres were 0-for-6 with the man advantage.
Buffalo outshot Chicago 41-21 in the game.
Chicago looks to continue their winning ways next game with a battle against the Blues. The Sabres will aim to get back on the winning track at Atlanta.


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Taste of Country Music Festival is the Greatest Lineup of the Summer
Normally you would have to travel down to Texas or Tennessee to see a festival lineup like the one coming to Hunter Mountain in June.
After many long hours and a great deal of work, some of country music biggest and brightest stars will be playing at the Hawk's 'Taste Of Country Music Festival,' the biggest and best, not to mention, the only 3-day country music festival in the Northeast, June 13-15 in Hunter, New York.
We are thrilled that Lady Antebellum will be headlining our first ever 'Taste Of Country Music Festival.' Since Hillary Scott is expecting, this is the only Summer show she committed to this Summer. Cue in "it's a quarter after one…"
When you think legends, the name Willie Nelson most likely would be on the top of your list. Willie Nelson and Family have been known to play as long as the fans want them to play. So you know you'll get your money's worth with Willie.
Fresh off NBC's 'Celebrity Apprentice,' Trace Adkins will be sharing the many hits of his 13 year career with his country-rock, country blues sty-lings. ,
Billy Currington sings 'Pretty Good At Drinking Beer,' but he's also pretty good at entertaining his many fans and is sure to get more than a few ladies excited.
Hunter Hayes is the hottest new act in country music. Hunter has the distinction of writing, producing, lead locals as well as all the background vocals and most of the instruments on his smash debut self titled album.
Montgomery Gentry will be rockin' the crowd with their unique style.
Joe Nichols was a favorite when he played Binghamton is looking forward to playing for Hawk fans again.
And Gloriana, Justin Moore, Dustin Lynch, Blackberry Smoke are some of the hottest new acts in country music today. We're thrilled to have their energy as part of the big weekend.
The show is going down June 13-15, 2013 at Hunter Mountain in the gorgeous Catskills.
This will be the concert everyone will be talking about and tickets can be purchased at tasteofcountryfestival.com or by clicking HERE There are also some great camping packages available.
The event of the Summer. Everyone will be talking about the 'Taste of Country Music Festival' June 13-15th under the huge tents at Hunter Mountain Resort in Hunter, New York.

subset: commoncrawl


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The National Transportation Safety Board's preliminary report on the Ocean City plane crash that killed two off-duty police officers says the plane appeared intact as it hit the ocean and it was found together, but fractured from impact.
The report states that divers found the fractured small military-style plane together on the ocean floor but the left wing was lost during recovery. The NTSB says investigators hope to salvage video from a camera in the rear cockpit. A second report will be released when the investigation is complete and a final report will include findings and a probable cause for the crash.
Forty-three-year-old Thomas Geoghegan Jr. and 27-year-old Joshua Adickes of Berlin were killed June 30 when the plane crashed shortly after takeoff from the Ocean City Municipal Airport.
PREVIOUS COVERAGE ON NBCWashington.com:
- Funerals Scheduled for Ocean City Plane Crash Victims
- Small Plane Crashes Into Ocean in Maryland
- Victims of Ocean City Plane Crash Identified as Police Officers
- Crews Removing Wreckage of Crashed Plane in Ocean City


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The first step to improve productivity is to recognise what it is, why it needs to be measured, and how to measure it.
A lack of definition and understanding of productivity is a common problem in most organisations. It can mean different things depending on what part of the business someone is from. But a definition is actually simple: it is an efficiency measure.
However, it should not be used as the only measurement in an organisation.
A good balance of these measurement criteria are needed with a sharper focus on efficiency.
There are four common reasons why productivity should be measured:
When organisations are ready to measure productivity, they should ask:
<< Previous
EY refers to the global organization, and may refer to one or more, of the member firms of Ernst & Young Global Limited, each of which is a separate legal entity. Ernst & Young Global Limited, a UK company limited by guarantee, does not provide services to clients.


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A Question About a Saying by Athanasius
BY Mark Shea
| Posted 11/28/11 at 1:00 AM
A reader asks:
How would explain to a critic of the Catholic Church CCC460, which quotes Athanasius about "becoming God"
I understand that it means "participation in the divine nature" but how would explain the strong language used? Poetic vs. technical theology?
I'd go with poetic emphasis. Obviously we are not Mormons and don't believe we are transformed into a God (a fact Athanasius knew very well). Nor do we believe that God the Father was once human and then graduated to godhood. Nor are we polytheists who believe the persons of the Trinity are three gods. Athanasius' point was indeed to drive home the reality that, as 2 Peter 1:4 teaches, we become divinized in Christ ("partakers in the divine nature), yet remain ourselves.
That passage from Athanasius, by the way, is a favorite with Mormon apologists (i.e, "God became man that man might become God.") It gets trotted out as "proof" that early Christians were basically Mormon before the Great Apostasy (when the wicked Catholics drove the true Church into oblivion, necessitating the re-revelation given to Joseph Smith as a patchup on the botched job Jesus and the apostles did.
The problem: For Mormonism to work, the Great Apostasy has to have happened centuries before Athanasius (who lives over 300 years after the Church supposedly went off the rails). Because by his time everybody (including Athanasius) looks, acts, talks, walks and quacks exactly like a Catholic and believes all sorts of things no Mormon would touch with a barge pole. That's why that quote from Athanasius, ripped bleeding from its context, is the only thing Mormon apologists know about him. Of his orthodox (not Mormon polytheistic) Trinitarianism, his devotion to the teaching of Nicaea, his faith in the Real Presence, his devotion to the Blessed Virgin as sinless and ever-virgin Mother of God and all the rest that marks him out as an "apostate" Catholic (by Mormon lights) and not a Mormon in the slightest, Mormon apologists say nothing.
My recommendation for chatting with Mormons: Don't waste time talking about the staggeringly huge doctrinal difference between Catholic faith and the King James Version fan fiction known as the Book of Mormon. Instead, ask for the exact date of the Great Apostasy. Because Mormonism has no other raison d'etre in the universe than as a remedy for that alleged Great Apostasy. If there was no Great Apostasy (which, you know, there wasn't) there is absolutely no point to Mormonism. It becomes a cure for a non-existent disease peddled by a 19th Century snake oil salesman who sold his wares to people who knew nothing about the history of the Church.
Copyright © 2013 EWTN News, Inc. All rights reserved.


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Here are the interesting links I came across on my travels around the web this week! Enjoy and have a great weekend!
- 25 Things you Can Re-use at Home
- 5 Home Remedies for a Sore Throat
- 10 Ridiculously Easy Fall Projects
- Choosing Cheerfulness
- How to Stop Spending Money
- Honey Chai Flavoured Roasted Almonds
- DIY Wordy Hairpins
- 10 Minute Super Hero Costume


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In a move so devious it might actually elicit sincere admiration, an ex-boyfriend of one Jennifer Fitzgerald used the Chicago airport to get back at her.
Apparently Brandon Preveau, Jennifer's ex, bought a an old used car in her name and parked it at the airport, where it has been piling up violations and tickets since around 2009. Now, Jennifer owes over $100,000 – the largest parking fine Chicago has ever seen.
Salt expresses how impressed he is with this plan, and he, along with Karson, share stories of similar revenge plots that they have played parts in over the years.
Click Below To Listen To Some Wicked Stories Of Revenge:


text: Hey guys I've always hated this. I am not on a data plan, and whenever I turn my phone on or randomly every once in a while the stock weather app says weather service unavailable because I have no connection, this is fine i Don't want connection cause of not having a data plan. So I Kill the weather app but it just keeps coming back up to run, any ideas on how to keep it killed?


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Europe in crisis, and the lessons of the Thirties
Few of us these days can recall the Thirties, but everyone can read about them and see that Europe was then politically sick.
Hitler ruled in Germany, Mussolini in Italy. General Franco won in Spain, General Metaxas in Greece. Austria succumbed to the 'Austrofascism' of Chancellor Engelbert Dollfuss. The relatively benign Dr Salazar ruled with a firm hand in Portugal. Admiral Horthy, an admiral without a ship, held power in Hungary, a nation without a navy.
The countries had all fallen a long way from the supposed, bright democratic future which the Versailles conference had laid out after World War I.
Fascism: The rise of extremists such as Mussolini and Hitler was a result of the economic depression of the Thirties
By the late Thirties, this whole panorama of democracy had been swept away by the economic depression of that decade. The dictators were a response to the belief that a serious crisis demanded strong men and stern measures.
To think that Europe could head off again in that direction may seem absurd. Surely the democratic system is entrenched? They said that last time.
Extremist parties are on the rise in Continental Europe, as recent elections have shown. In Greece, the far-Left party Syriza is reckoned to have an evens chance of winning at the forthcoming polls. At the last and inconclusive election the far-Right Golden Dawn, openly fascist, won 21 seats.
Far-right: Greece's Golden Dawn party gained 21 seats in that country's elections
In France, Marine Le Pen's National Front had its best ever result. Immigration is an issue throughout Europe. Note, too, that in both Greece and Italy the premiers have been appointed precisely because they are not elected politicians but 'technocrats', a fancy name for bureaucrats.
Before you ask — before it writes me angry letters — note that UKIP is not an extremist party, whatever the sneers of David Cameron. However, it adds substantially to the swing away from normal voting patterns.
A survey by thinktank Demos finds that the young in Europe are abandoning orthodox voting patterns. Expect this to continue as long as the euro-crisis goes on — as it will because no one seems able to make up their mind about what to do next.
In France, Marine Le Pen's far-right National Front has gained huge support
Various proposals are made for solving the problem of the peripheral governments which cannot borrow on the open market and — they say by coincidence — with banks badly in need of rescues.
Both are merely symptoms of having spent and lent too much, some of it in the quite recent past. The blame for not dealing with these issues earlier attaches to the banks and governments themselves.
Who thought the Spanish property boom would go on for ever? Who really believed that the Greek government would repair years of mismanagement by borrowing even more from its eurozone neighbours?
During this protracted crisis, we have all those grand figures in politics — and supposedly learned figures in economics — insisting for two years now that Greece would be rescued and the euro was safe.
The plans being tossed around at the moment, Europe-wide bonds, a single Eurozone ministry of finance, a rescue of banks by using the funds reserved for rescuing governments — all suffer from the weakness that they depend on Germans underwriting potentially astronomic losses.
The Germans do not wish to do so. They cannot see why they should prop up the whole system because they have worked harder, borrowed less and prospered more.
More from Andrew Alexander for the Daily Mail...
- ANDREW ALEXANDER: Rebels, ploys and a matter of life and death 14/01/14
- Is it poisoned chalice time for Theresa? 10/12/13
- ANDREW ALEXANDER: Did David Cameron see the truth in Sri Lanka? 20/11/13
- ANDREW ALEXANDER: Why the EU is desperate for Britain to stay 05/11/13
- Oh no, the U.S. has dropped us in it again 29/10/13
- The 'beauty' of VAT? A vicious circle that turns us all into losers, by ANDREW ALEXANDER 15/10/13
- ANDREW ALEXANDER: Beware the Tory without ambition... 01/10/13
- Don't be naive... politics is nasty, not nice 24/09/13
- After the blow struck for democracy last week, at last, the U.S. will be without its usual lap dog 03/09/13
Supposedly a crushing argument is that if Greece left the euro there would be bank runs. I have news for those intimidated by that fear.
The 'runs' have already happened. Huge sums have been transferred from eurozone nations by the mere pressing of buttons on the key pad.
We are in a state where extremist parties can gain real power. Even if not outright winners, they may hold the balance of power (as in Hitler's case). The EU dream of ever-rising amity and prosperity remains a costly fantasy.
'Official' spending? We're paying!
We are told from all sides that austerity has failed. We should have less of it and (somehow) spend more money that we do not have.
My favourite approach to this is voiced in the City by Terry Smith, of brokers Tullett Prebon, whose firm reckons that Government spending has risen in real terms, ie, allowing for inflation, by 50 per cent over the past ten years — and back then we seemed happy enough. What is it that we have now which we could not possibly do without? Answer: nothing.
But you would not get that impression from TV and radio, which forms so many people's political views. There comes along, say, the news that the Government is to cut £x million from . . . choose any service you like.
Surely, comes the interviewer's question, this will have a serious impact on the poor/the (hard- working) middle class/the kiddies/ immigrants/the North-East/the South-West/pensioners/those not yet pensioners etc.
This is, of course, a leading question with the implication that official spending is inherently a good thing.
Say at some point that this is taxpayers' (hard-won) money and you might have a reasoned debate. Too much to ask?


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catastrophic wheel failure - why?
Look at the photos below of what happened to my wheel today:
Facts are as follows:
This wheel was 36-spoke, 3-cross, custom built, tensioned, and true.
The wheel had about 500 miles on it when it detonated.
The tire was a WTB Slickisaurus 700x37c inflated to (Mfg. recommended) 85 psig.
The tube showed about a 9-inch-long smooth split on the spoke-side of the tube
The rim tape was not punctured
The wheel detonated with a bang on smooth pavement at about 19 mph
The rim did NOT fail at the weld - the weld is still intact.
The wheel bead showed no signs of damage other than where it was punctured by the broken rim
The LBS does NOT think this was caused by the bead coming loose from the rim (their opinion)
What caused this rim to fail?
What can I do to prevent this from happening again? (I'm lucky to be alive)
Does Mavic owe me another wheel?
I have three other wheels with the same rims - am I at risk from them too?
Missing mucho info.
What were you doing when the wheel broke?
What did you hit or what hit you?
Why does the wheel appear to be too big for the bike?
Why are you riding around without a seat?
What were you doing when the wheel broke?
I was riding on smooth pavement at 19mph. I was about 40 minutes into my daily ride. No potholes, no debris - smooth pavement.
What did you hit or what hit you?
No collision whatsoever. One moment everything was normal, the next, I was sliding down the road on my butt. I believe when the wheel detonated, I was thrown up. When I came down, the seatpost bolt broke and dumped me off the back of the bike. Luckily, I didn't land on the seatpost or the broken rim or I'd not be here now.
Why does the wheel appear to be too big for the bike?
Must be an artifact of the photo - the bike is a Klein Navigator touring frame made for (and with clearance for) the 700x37c tires that are on the bike.
IED maybe? Did you also draw sniper fire?:D
Seriously, boyo, you had enough good luck left to walk away.
I find the secondary faliure quite interesting, a fracture caused by the rapid de-tensioning of the rim.
Interesting, as if your rim sheared in half once the tire-tube unit stopped rotating at the same rate as the rim and hub. That rim is a monocoque thin-wall design, not a "boxed" design, right?
poorly built wheel?
The position of the tear in the rim makes sense--8 and 2 o'clock--which is close to the horizontal direction of movement of the bike with a slightly nose-down attitude, typical for the nose-down dive that takes place in an emergency braking, and with the center of mass--mostly you--above the line of shear. Did you leave rubber on the pavement?
The position of the tear in the rim makes sense--8 and 2 o'clock--which is close to the horizontal direction of movement of the bike with a slightly nose-down attitude, typical for the nose-down dive that takes place in an emergency braking, and with the center of mass--mostly you--above the line of shear. Did you leave rubber on the pavement?
who built this timebomb? if your LBS did and it hasnt been too long since you've got it, they shouldn't give you much trouble. Mavic may have had some machining error, but I might try to get the wheelbuilder to give you a new one. you could have been seriously hurt.
No spokes broke? None were pulled through the rim? I wonder if it's not just a defective rim from Mavic.
I don't have a lot of experience with thin-wall rims (if that is indeed what you have), but 85 PSI doesn't seem to be enough pressure for that kind of damage at the primary break. The clean break was incured my the sudden stress loading that the first break caused in what remained (for milliseconds) a tensioned wheel.
If the tube went, the metal damage on the rim would be blown OUT, not folded inward as shown in the pics.
To quote one of my trainning offcers may years ago, "sure beats the hell out of me!"
A guess...
Maybe the wheel was stress-relieved by bending it, while holding on to the rim, and the builder used too much force, cracking the rim in the middle - i.e. in two places diametrically opposite.
Did the chain break? The rear part of the drivetrain doesn't look quite right.
I have no idea what could have caused that but, I'm glad you're OK.
Never seen anything like it.
I don't have any answers but am glad you are ok. It looks horrible! Yikes!
Looking at all the parts of the wheel there are certain places you can rule out: hub, spokes (none broke, tire (you mentioned no damage to the tire, right?), rim tape is intact.
Only the tube and the rim are damaged (and considerably).
HOLY CRAP! :eek:
looking at the evidence the only conclusion i can come up with is that the rim was defective, that or it was tensioned way too high - not to put down your lbs.
I've thought and thought about this one. I'd think you can pretty much rule out a material problem because of the dual failure. The only thing that makes sense to me is a poor wheel build. If a number of spokes 180 deg opposite from each other were way too tight, and the ones 90 deg out from these were then too loose it would cause rim forces much like if you were to push down on a hula hoop. Every revolution of the wheel would be the equivalent of pushing and letting up on the hoop. The aluminum would fatigue pretty quickly at the two sides where the spokes were loose. When the first side failed catastrophicaly that would send a shock wave around the rim causing the failure at the opposite side.
Mavic has had quaility control problems as of late. My LBS is contemplating dropping them.
This wheel has failed as I would have expected it to if it had an explosive decompression near the valve.
Look carefully. The wheel is in about the same position as when the accident occurred. Decompression near the valve (on the road at this time) causes a rapid destressing of the rim. The rim "springs out", and the elastic shock propogates (via the hub) to the top spokes (near the brake), pulling the rim down at the top. See how the top of the rim has been flattened, and at least one spoke head (near the label) is partially pulled out of the rim. The top spokes are already loaded at this time by the weight of the rider - remember, the bike hangs off the top spokes. The deformation (flattening) of the top half of the rim causes the rim to bulge out in the for/aft positions, ie. to make the rim elliptical. It is likely that at this time, the bottom half of the rim has elastically rebounded and is therefore contributing to the fore/aft elongation.
The result of this is a classical shear failure at the fore/aft positions as shown in the photo. We would expect a shear failure would be at about 45 degrees, which is exactly what we see. The initiator for the shear failures is, as expected, spoke holes, which are the local stress raisers.
I see no pre-existing contributor to this failure. It's just plain bad luck. You got an explosive blowout just at the point that the wheel contacted the ground and all the other dynamics were just not in your favour - saddle bouncing, slight road irregularities, pedal torque - it all came together at the wrong time.
I could be wrong and I only have the supplied photo to work from, however I am an engineer and have done this stuff before. Any other engineers care to comment on this failure - an ATSB accident investigator would be just dandy!!!
I am interested in what other have to say about this failure. I ride on Mavic wheels (MA40's and M193's), weigh 95kg, and have no interest in a set of circumstances that may result in the seatpost comming out my nostrils!!!
hope this helps,
I've had another close look at the photo. The left hand side of the wheel certainly failed in sheer as expected. Clearly, the subsequent collapse of the wheel cause the failure of the right hand side of the wheel, which appears on closer inspection to have a brittle bending failure. See how the rim appears to have opened up from the outer side, the fracture propogating back towards the nipple hole. On the other side, a crack has run along the rim terminating in a shear failure out to the edge, just as expected for a failure in bending.
I think there is some possibility that this rim was overly brittle. How much does the rider weigh and how many miles on the wheel?? I would get the wheel to the metallurgical department of the local Uni and have it examined.
I note that there are no ferrules in this rim. Each to his own, but I will not ride a rim without ferrules, or some other equivalent destressing arrangement.
Three important issues not discussed here:
1. FarHorizon, how heavy are you and is there any specifications for maximum wheel weight?
2. Did you contact directly Mavic in the country where you live? if not, do it, Mavic engineers and developers would be most interested on checking that wheel, and certainly will replace you the wheel.
3. Was the tire width size within the limits for the rim specification?
Looks to me as if the wheel was dropped from the top of a 10 story building before it was installed on the frame.
|All times are GMT -6. The time now is 09:50 AM.


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USDA's plans to forge a consulting contract with former Agriculture Department official Shirley Sherrod have turned rocky. Sherrod, who met briefly with USDA Secretary Tom Vilsack and his staff two weeks ago about contracting with USDA to help the department address long-standing civil rights issues, has said she was insulted by the department's initial offer.
Vilsack fired Sherrod last year after media reports misconstrued her statements about white farmers as racially insensitive. She was offered a contract that her spokesman valued at about $35,000 — without expenses for travel.
"The resources that would be available for her don't work for the things that they are proposing," said Sherrod's spokesman Drew Berry. "She is not angry with Vilsack, but she is not happy with the process and what has transpired. She is willing to help, but she is not going to do anything in a superficial way."
USDA spokesman Justin DeJong said "discussions are ongoing," and department officials told members of the Congressional Black Caucus — who took up the issue of Sherrod's contract at their weekly meeting Wednesday — that the issue is a misunderstanding.
Berry said that Sherrod, who was scheduled to speak Thursday at a civil rights conference in North Carolina, had been mailed a copy of the contract this week. She was disappointed and found it insufficient for the scope of the work. DeJong said the "contract that was sent to her was a draft." The consulting work would not be a full-time position.
On Wednesday, Sherrod told American Urban Radio Networks that "talks had broken down," but Berry and USDA officials agreed that the negotiations are ongoing.
USDA officials have said the potential contract with Sherrod, who was ousted last summer in a racial imbroglio that embarrassed the department, is a part of its plan to fix long-standing problems with discrimination.
The USDA is also seeking contracts with the National Latino Farmers & Ranchers Trade Association and the Intertribal Agriculture Council. Those groups and Sherrod's would be tasked with implementing reforms recommended in a two-year, $8 million civil rights assessment released last week by the department.
After reports surfaced last week that Sherrod would return to USDA, Sherrod said she was upset that the information had leaked and considered any announcement that she would return to USDA "premature."


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Facebook is testing a feature for its Android users that syncs photos taken on their mobile devices directly to their Facebook accounts.
The social network confirmed the test to The Verge, saying it was on a "really small" scale, and not offering further details on whether it would be rolled out to more users.
The Verge described the photo sync feature as a hybrid of the instant photo upload feature on Google Plus, where photos are automatically uploaded to users' accounts, but to a private folder, and Apple's Photostream, which uploads photos to the cloud and syncs them with all of a user's iOS devices.
According to The Verge, users can determine if they have access to photo sync by going to their timelines on Facebook for Android, tapping photos, and tapping the sync button if it appears.
Readers on Android: Do any of you have access to photo sync?


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According to TMZ.com, two tour buses that are being used on the "The Soundtrack Of Summer" trek featuring STYX and FOREIGNER went up in flames early Wednesday afternoon in Philadelphia. The fire started shortly after noon at a tour bus parking facility and involved only "crew buses" and not "band buses," according to the report.The tour buses were reportedly empty and there were no reports of any injuries. The exact cause of the fire is still under investigation. The bands are scheduled to perform together Thursday, July 3 at Susquehanna Bank Center in Camden, New Jersey. STYX, FOREIGNER and former EAGLES guitarist Don Felder kicked off their three-month tour on May 16 in Oklahoma City. STYX and FOREIGNER previously toured together as part of the 2010 "Union" trek.
STYX And FOREIGNER 'The Soundtrack Of Summer' Tour Buses Go Up In Flames
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The Big Grape Event
Grand Island Hy-Vee has announced plans for the 2013 "Big Grape" event to be held October 19, 2013 at the Pinnacle Bank Arena at Fonner Park. The event, attended by over 800 guests last year, is a premier wine & food experience that raises money for the Juvenile Diabetes Research Foundation. Guests this year can once again look forward to live & silent auctions, hundreds of wine samples and an array of gourmet foods to try throughout the evening.
The Big Grape 2012 raised over $68,000 the night of the event and all totaled, the group donated over $71,000 to JDRF. This made the Grand Island Hy-Vee the number one store for JDRF fundraising in the entire company.
"We couldn't be more proud of how our community rallies around this fundraiser," Dave Blum, Store Director of the Grand Island Hy-Vee says. "We were the number one fundraiser out of all of the Hy-Vee stores. It is a terrific accomplishment but more importantly the "sibling rivalry" throughout Hy-Vee land challenges us to raise even more money for JDRF. We never lose sight of our mission; it's for the kids after all. But winning is nice too."
Tickets are now available at Hy-Vee's customer service counter. Tickets are $50 in advance or $60 at the door. "The Big Grape" committee is currently obtaining sponsorships for this year's event. For more information on how to become a sponsor or how your business can be a part of "The Big Grape", you may contact Ken Pedersen at Hy-Vee, 308-381-3678 or by email email@example.com.


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Volunteer Opportunities
At the Blues Festival
The Mimbres Region Arts Council is dependent on its strong volunteer base to support a diversity of cultural events for our region. Whether it's taking tickets at shows, designing murals with students, making gourmet chocolates for Chocolate Fantasia, or helping to produce music festivals, there is something for everyone!
Contact Elizabeth at email@example.com to learn more.


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Protein traffic is necessary to maintain homeostasis in all eukaryotic organisms. All newly synthesized secretory proteins destined to the secretory and endolysosmal systems are transported from the endoplasmic reticulum to the Golgi before delivery to their final destinations. Here, we describe the COPII and COPI coating machineries that generate carrier vesicles and the tethers and SNAREs that mediate COPII and COPI vesicle fusion at the ER-Golgi interface.
Secretory Compartments of the Endoplasmic Reticulum-Golgi Interface
In eukaryotic cells, secretion involves movement of cargoes through a linear assembly of membrane-bound compartments. The secretory pathway is comprised of structurally distinct organelles and membrane-bound transport intermediates that facilitate transport between them. The advent of live imaging and the identification of specific compartmental markers have changed the face of the secretory pathway. Currently, the compartments of the secretory pathway, beginning with the site of cargo synthesis, include the endoplasmic reticulum (ER), ER exit sites (ERES), the ER-to-Golgi intermediate compartment (ERGIC), the Golgi complex, and the trans-Golgi network (TGN) (FIGURE 1). In some cell types, endosomes are also traversed by cargo proteins and thus also are considered secretory compartments.
ER and ERES
All newly synthesized secretory and plasma membrane (PM) proteins and proteins that reside in the ERGIC, Golgi, endosomes, and lysosomes are translated on the cytoplasmic surface of the rough ER (RER) and are either inserted into RER membrane or translocated into the RER lumen. The advent of new organellar markers and live imaging of cargo movement established that proteins are exported from the RER only from specialized ERES (FIGURE 2A). ERES are morphologically defined as smooth projections of the ER that are coated with COPII coat components (see below). ERES give rise to COPII-coated vesicles. Mammalian cells generally have 50–70 ERES of ∼0.5-μm diameter randomly scattered along the ER with some concentration in an area adjacent to the Golgi (67, 165). ERES are relatively stable, persisting for >10 min. Thus these sites are likely to generate numerous COPII vesicles during their existence. ERES-generated COPII vesicles transport cargo to the next compartment in the secretory pathway, the ERGIC.
The ERGIC (also known as vesicular-tubular clusters or pre-Golgi intermediates) is a collective name given to pleiomorphic compartments composed of vesiculo-tubular profiles that are found adjacent to ERES (FIGURE 2B). Although the exact nature of the ERGIC is still a matter of debate, the ERGIC is distinct in biochemical composition from the ER and the Golgi (150) and is not continuous with either the ER or the Golgi (26). Individual ERGIC units seem to communicate laterally through rapidly forming and dissipating tubules and vesicles (17).
Live imaging of the ERGIC marker ERGIC53 shows that individual ERGIC are stable compartments that remain close to ERES, without net movement toward the Golgi (17). In contrast, live imaging of secretory cargo shows separation of large (typically 1 μm in diameter) anterograde-destined carriers containing cargo from ERGIC53-labeled stable ERGIC. Cargo-containing carriers track on microtubules toward the microtubule organizing center (MTOC) and the Golgi complex (70, 128). Thus it appears that ERGIC is a stationary sorting compartment that gives rise to relatively large anterograde-destined transport intermediates.
In addition to generating anterograde carriers for cargo transport to the Golgi, the ERGIC sorts proteins into at least two additional pathways: recycling proteins are sent back to the ER and resident ERGIC proteins are retained within the ERGIC. Sorting into the recycling pathway is most clearly understood and is mediated by the COPI coat, which facilitates formation of COPI vesicles (described in detail below). It is less clear whether COPI also participates in the formation of the large anterograde carriers and/or protein retention within the ERGIC. However, it is clear that COPI is recruited to the anterograde carriers as they translocate from the peripheral ERGIC toward the MTOC (166), suggesting that COPI-coated vesicles continuously bud from the carriers.
The Golgi complex exhibits distinct morphologies in different organisms. In mammals, the Golgi is organized into a single ribbon of stacked cisterna (50, 84). In most model organisms, including the worm C. elegans, the fly D. melanogaster, and the plant A. thaliana, the Golgi exist as discrete stacked elements (Golgi mini-stacks) scattered throughout the cell (51). In the yeast S. cerevisiae, the Golgi exist in discrete tubules scattered throughout the cell (2).
Irrespective of morphology, the Golgi in all higher eukaryotes is composed of closely packed membrane cisterna with a defined polarity, cis facing the ER, trans facing the opposite side, and medial in between (FIGURE 2C). The number of cisterna varies in different organisms and in distinct tissues, with most cells having four to seven cisterna. The polarity of the Golgi is most clearly reflected in the localization of various glycosyl-transferases. Enzymes such as the early acting mannosidase II are detected on the cis-side, whereas the late-acting galactosyl-transferase is detected in the trans-cisterna (95, 136, 176, 178, 183).
The Golgi complex is flanked on the cis-side by a tubular cis-Golgi network (CGN) and on the trans-side by the TGN. The CGN may consist of fused and concentrated transport intermediates approaching the Golgi from the ER. The relationship between the ERGIC, anterograde carriers, and the CGN is unclear. The current model is consistent with carriers originating from the ERGIC, tracking on microtubules toward the MTOC and fusing there with each other to generate a new cis-cisterna. The TGN consists of tubular elements that are distinct from the trans-Golgi cisterna. The TGN appears to be a complex compartment consisting of distinct subdomains that contain different coiled-coil proteins called golgins (43, 44, 62). From the TGN, transport intermediates can either traffic directly to the PM or can deliver cargo to the endosomal system or to secretory granules (FIGURE 1).
Transport through the Golgi has been extensively studied and even more extensively debated. Multiple models have been considered and the one gaining the most recognition is the cisternal maturation model (63, 127, 168, 188). In this model, anterograde carriers formed from ERGIC translocate toward the MTOC and accumulate at the cis-Golgi region where they fuse into a network that subsequently becomes the cis-Golgi cisterna. Newly formed cis-cisterna progressively move toward the trans-side as they remodel by losing early acting Golgi enzymes and acquiring late-acting ones (24). This model implies that the secretory cargo remains inside the original membrane compartment throughout transit, while the compartment itself undergoes complete remodeling through the loss of resident early Golgi proteins and the acquisition of late ones (167). Remodeling occurs through packaging of Golgi proteins into COPI vesicles that move in the retrograde direction from trans- to cis-Golgi cisterna. In this model, the full complement of Golgi resident proteins is continuously recycled to the more proximal compartment.
Increasing Complexity of the Trafficking Machinery
The last few decades have witnessed an explosive growth in defining distinct stages of traffic and identifying molecules that regulate each stage of transport. The functions of coat complexes, Sar/ARF GTPases and their activating guanine nucleotide exchange factors (GEFs), and deactivating GTPase activating proteins (GAPs), tethering factors, and soluble N-ethylmaleimide sensitive factor receptors (SNAREs) in membrane trafficking are clearly established (FIGURE 3). Below, we summarize the general understanding of this basic machinery responsible for cargo traffic at the ER-Golgi interface.
Vesicle Formation at the ER-Golgi Interface
COPII- and COPI-coated vesicles are operational at the ER-Golgi interface. The formation of both types of vesicles is mechanistically conserved and in both cases involves coat complexes, Sar1/ARF GTPases, their GEFs, and their GAPs.
Anterograde COPII vesicles.
Cargo sorting for exit from the ER occurs exclusively at ERES and is mediated by the COPII coat, which binds and concentrates secretory cargoes into vesicles (9, 19, 93, 142, 143, 177, 204). COPII recruitment is mediated by the active GTP-bound form of the small GTPase Sar1. Like all GTPases, Sar1 exhibits low intrinsic GDP-to-GTP exchange, and in cells this process is catalyzed by the Sec12 GEF (11, 143, 189). Sec12 is a membrane protein localized to the ER, which restricts Sar1 activation to the ER. Active Sar1 directly binds the Sec23/Sec24 heterodimeric subcomplex of the COPII coat, which in turn recruits the Sec13/Sec31 heterotetrameric subcomplex to the Sec23/Sec24 core. Sar1, Sec23/24, and Sec13/31 are the minimal components required to generate COPII vesicles in vitro (106). Sec12 is required in addition to reconstitute GTP-dependent COPII vesicle budding (59). There are two Sar1 paralogs (Sar1a and Sar1b), two Sec23 paralogs (Sec23a and Sec23b), four Sec24 paralogs (Sec24a–d), a single Sec13 paralog, and two Sec31 paralogs (Sec31a and Sec31b). The exact functional relationships between these components to make distinct COPII vesicles remain to be defined.
Sorting of transmembrane cargo proteins into COPII vesicles is mediated by the Sec24 subunit of COPII, and different isoforms of Sec24 have been shown to bind different sorting motifs and thus expand the range of cargos that can be exported out of the ER. Sec24 binds to the cytoplasmic tails of transmembrane cargo proteins and sequesters them into the nascent vesicles. Sec24 isoforms a and b (but not isoforms c and d) bind the DXE motif and also the LXXL/ME motif on cargo proteins. Sec24 isoforms c and d bind the IXM motif (101).
Soluble cargo proteins within the ER lumen are exported from the ER by binding to cargo receptors of the ERGIC-53 family, the p24 family, or the Erv family that facilitate their sorting into COPII vesicles. These transmembrane receptors contain lumen-exposed domains that bind cargo proteins and cytoplasmic domains that interact with COPII components for inclusion into COPII vesicles.
ERGIC-53 is a single-pass transmembrane protein with a large NH2-terminal luminal domain that binds glycosylated cargo proteins (6, 82). Additional members of the ERGIC-53 family include ERGL, VIP36, and VIPL that display distinct binding preferences for high mannose containing glycoproteins (80). The function of these receptors as cargo transporters implies that they bind cargo within the ER and release it within the ERGIC/Golgi before recycling back to the ER. The regulation of cargo binding and release appears to be mediated by a decrease in intraluminal pH between the ER (∼7.2) and the Golgi (∼6.4), decreased Ca2+ levels, and possibly phosphorylation (68, 169).
The p24 family members are similar to ERGIC-53 and contain a luminal NH2-terminus, a single transmembrane segment, and a small cytoplasmic tail that binds to COPII components (169). The p24 proteins are ubiquitous and, in mammals, consist of 8–10 isoforms with distinct expression patterns and possibly different cargo binding preferences (170).
The Erv family members are multi-spanning transmembrane proteins with varying topologies (reviewed in Ref. 41). The types of cargoes they bind vary, and Erv29 appears to bind preferentially soluble proteins, whereas both Erv14 and Erv26 appear to traffic preferentially transmembrane proteins. A recent review provides more detail on the function of the ERGIC-53, p14, and Erv classes of cargo receptors (41).
The critical roles of cargo receptors are underscored by diseases associated with mutations in various receptors. Mutations in ERGIC-53 cause autosomal recessive bleeding disorders due to defects in trafficking of coagulation factors V and VI (205, 206). Deletion of only the p23 member of the p24 family in mice causes embryonic lethality (42). Interestingly, deletion of all eight members of the p24 family in yeast maintains cell viability despite trafficking defects of select cargoes (16, 145). Clearly, the trafficking of essential cargoes by distinct p24 proteins is essential for metazoan development, even if the removal of all p24 proteins can be tolerated in a single eukaryote cell grown under laboratory conditions.
Importantly, all the cargo receptors in the ERGIC-53, p24, and Erv families continuously cycle between the ER and the ERGIC/Golgi. All contain cytoplasmically exposed motifs for interactions with COPII components for inclusion into COPII vesicles, as well as sorting motifs recognized by COPI components for inclusion into recycling COPI vesicles.
In contrast to the cargo receptors outlined above that are packaged into COPII vesicles, additional factors that do not enter COPII vesicles participate in traffic of specific cargoes, such as the large, >300 nm in length fibrillar collagens. TANGO1 (transport and Golgi organization) factor (also called melanoma inhibitory activity member 3 or MIA3) is a transmembrane protein localized to ERES that was initially shown to be required for efficient exit of collagen VII from the ER (139). TANGO1 binds collagen VII within the ER lumen and links it to the Sec23/24 subunits of COPII within the cytosol and thus loads collagen VII into nascent COPII vesicles. The essential function of TANGO1 is underscored by the severe chondrodysplasia and death at birth of homozygous TANGO1-null mice (193). TANGO-null mice show defects in trafficking of all collagens examined (I, II, III, IV, VII and IX), but traffic of other components of extracellular matrix, such as fibronectin or aggrecan, is not impaired, indicating that TANGO1 is a cargo-selective "loader." Recently, TANGO has been shown to interact with cTAGE5 (cutaneous T-cell lymphoma associated antigen 5), a transmembrane protein localized to ERES (140). Like TANGO1, cTAGE5 binds Sec23/24 subunits of COPII and is required for efficient secretion of collagen VII. Thus cTAGE5 might be a "co-loader" of collagen into COPII vesicles. Neither TANGO nor cTAGE5 homologs are detected in yeast, suggesting that they do not represent the core COPII vesicle budding machinery.
Sorting of cargo into COPII vesicles is also regulated by the Sec23 subunit, which acts as a GAP for Sar1 and may proofread cargo sequestration into nascent vesicles (19). Sar1 GTPase activity is further stimulated by binding of Sec13/Sec31, which may ensure that complete COPII coats only assemble after sorting of cargoes into the nascent vesicle. GAP activity may also participate in the eventual disassembly of the COPII coat, although the details of uncoating remain under investigation.
Membrane deformation leading to the formation of COPII vesicles is likely facilitated by Sar1 and the Sec23/24 complex and then globally stabilized by the Sec13/31 complex. Sar1 has been shown to deform membranes in vitro (87) and has been suggested to facilitate vesicle scission in a manner analogous to that of dynamin, which mediates scission of clathrin-coated vesicles (7, 129). Vesicle separation requires the amphiphatic NH2-terminal helix of Sar1 that inserts into the membrane and initiates membrane constriction (21, 87). The size of COPII vesicles varies and in vivo appears to range from 50 to 90 nm. The size is probably regulated by the Sec13/Sec31 subcomplex because Sec13/Sec31 heterotetramers alone can self assemble into various size cages (58). The potential to form different sizes of COPII vesicles may be important for accommodating different size cargo that needs to exit the ER.
The formation of ERES and packaging of cargo into COPII vesicles is regulated by additional factors. Depletion or inactivation of the peripheral membrane protein Sec16 causes a decrease in the recruitment of COPII components to ERES and causes cargo accumulation within the ER (74, 75, 171). Sec16 has been shown to directly bind all four subunits of the COPII coat and is believed to stabilize COPII on membrane during COPII vesicle formation (48, 61, 152, 171). Sec16 targets to ERES independently of COPII (74). Sec16 appears to function together with its direct interactor TFG1 (TRK-fused gene) (194). TFG1 also localizes to ERES, and its targeting depends on Sec16 since depletion of Sec16 inhibits TFG1 recruitment to ERES. Depletion of TFG1 causes cargo accumulation within the ER (194). Both Sec16 and TFG1 are general traffic facilitators and are likely to influence the trafficking of multiple if not all cargo out of the ER. Their function is fundamentally different from that of TANGO and cTAGE5, which are cargo-specific regulators of trafficking.
Another factor that may function to stabilize the COPII coat is p125A, a soluble protein that is stably associated with the Sec13/Sec31 heterotetramer in the cytosol (121). P125A also binds Sec23, but this interaction appears to occur exclusively on the membrane. p125A is enriched at ERES, and depletion of p125A does not block traffic but causes delays in ER export of multiple cargos. Thus p125A has been proposed to stabilize the binding of the Sec13/Sec31 outer coat to the sec23/Sec24 inner coat and may have a general role in regulating COPII function.
Additional factors that control ERES structure and COPII vesicle formation include PCTAIRE kinase (126), p38 MAP kinase (185), phospatidylinositol (155), phosphatidylinositol-4 kinases (23), and the calcium binding ALG-2 (18). However, the mechanisms through which these components regulate ERES and COPII vesicle formation remain under investigation.
The integrity of the COPII coating machinery is essential for proper development and physiology. Loss of the Sar1b isoform expression or function causes accumulation of large (200–400 nm) chylomicrons within the ER (157). Mutations in the SEC23A gene cause cranio-lenticulo-sutural dysplasia due to defects in packaging of collagens into COPII vesicles (57, 179). In contrast, mutations in the SEC23B gene cause congenital dyserythropoietic anemia type II, an autosomal recessive disease probably due to incorrect glycosylation or trafficking of components required for erythropoiesis (20, 149). The distinct phenotypes caused by mutations in the highly related Sec23a and Sec23b underscore the high level of specialization of the COPII export machinery for specific cargoes in specific cell types. Similar to the loss of Sec23a in humans, defects in Sec24d cause skeletal defects in zebra fish due to inhibition in the trafficking of type II collagen and matrilin (119, 141).
Retrograde COPI vesicles.
COPI appears to be required at multiple stages of ER-to-Golgi transport. Initially, COPI was proposed to form vesicles for anterograde (forward) cargo transport (8, 122). However, subsequent studies implicated COPI in retrograde (from Golgi back to ER, and intra-Golgi) retrieval of proteins (90, 154, 164). In the currently accepted model, COPI vesicles may form from the ERGIC, from anterograde carriers as they move toward the Golgi, and from the Golgi cisterna. Such COPI vesicles may traffic all the way back to the ER or may transport cargo to the more proximal compartments (from trans- to medial-Golgi, from medial- to cis-Golgi, from cis-Golgi to ERGIC, and from ERGIC to ER).
The COPI coat is composed of heptameric (α, β, β′, γ, δ, ε, ζ) complexes called coatomers composed of two subcomplexes: a trimeric complex composed of α, β′, ε, and a tetrameric complex composed of γ, δ, ζ, and β (49, 187). Structural analysis of the α, β′, ε trimeric complex shows that it contains motifs similar to those in the Sec13/Sec31 complex that forms the outer layer of COPII-coated vesicles (86). Thus, the α, β′, ε trimeric complex is proposed to form the outer layer of the COPI coat. This is also supported by the findings that the γ COP subunit has structural similarity to α and β adaptin proteins (71) that form the inner core of the clathrin coat. Thus the present understanding is that the γ, δ, ζ, β tetrameric complex forms the inner core and that the α, β′, ε trimeric complex forms the outer layer of the COPI coat.
Two distinct isoforms of the γ (γ1 and γ2) and ζ (ζ1 and ζ2) subunits have been identified (22). The isoforms assemble into heptamers with the other five subunits, and analyses of coatomers in 3T3 cells indicate that ∼50% of coatomers contain the γ1ζ1 isoforms, ∼30% contain γ2ζ1, ∼20% contain γ1ζ2, and <5% contain γ2ζ2 (14).
Significantly, the different γ and ζ isoforms (and by extension the COPI coats containing these proteins) show distinct subcellular localization (109). The majority (∼70%) of COPI coats containing γ1 are localized within the cis-Golgi, whereas the majority (>60%) of COPI coats containing γ2 are found in the trans-Golgi. The majority (∼80%) of COPI coats containing ζ2 are found in the cis-Golgi. The localization of ζ1-containing COPI coats has not been determined as yet due to the lack of isoform-specific antibodies. This differential distribution of γ and ζ subunits occurs within the context of almost equal distribution of the β subunit, which shows 55% localization to the cis-Golgi and 45% to the trans-Golgi (109). This raises the key question of whether the different COPI coats serve different transport routes and traffic different cargo between different compartments.
Structurally and functionally distinct COPI vesicles have been described by numerous immunoelectron microscopic studies. Analysis of COPI vesicles in the scale-producing alga Scherffelia dubia shows two types of COPI vesicles: those with a light interior and a thin ∼11-nm coat, and those with darker content and a thicker ∼18-nm coat composed of an inner layer and an outer diffuse halo (47). Significantly, the "thin" COPI vesicles are found exclusively in the proximity of the cis-Golgi and in the space between the cis-Golgi and the ERES, suggesting that they recycle materials from the cis-Golgi to the ER. In contrast, the "thick" COPI vesicles localize exclusively adjacent to medial- and trans-Golgi, suggesting that they mediate retrograde trafficking within the Golgi. Only the "thick" COPI vesicles contain the Golgi enzymes sialyl-transferase and mannosidase I. The implication of this finding is that COPI recycling to the ER occurs exclusively from the cis-Golgi, never from medial- or trans-Golgi.
The existence of a population of COPI vesicles that shuttles from the Golgi to the ER that is distinct from intra-Golgi COPI vesicles is also apparent in mammalian cells because immunoelectron microscopy shows that COPI vesicles containing the KDEL receptor are distinct from COPI vesicles carrying the Golgi SNARE GOS28 (122). The separation of ER-destined COPI vesicles from intra-Golgi COPI vesicles is also supported by in vitro assays reconstituting COPI vesicle budding that show the formation of a population of COPI vesicles containing p24 family members p24α2, p24β1, p24δ1, and p24γ3 (ER-destined vesicles) and a population of COPI vesicles containing mannosidase II and the GOS28 SNARE (intra-Golgi vesicles) (85). The packaging of p24 proteins into COPI vesicles that are separate from COPI vesicles that carry mannosidase II has been confirmed in another study (100). These two populations of COPI vesicles could be further distinguished by their different tethering specificity (discussed below).
The involvement of COPI vesicles in anterograde trafficking, although currently out of vogue, is supported by experimental evidence. Immunoelectron microscopy showed COPI vesicles containing the anterograde cargos proinsulin and the viral glycoprotein VSV-G (122, 123). However, analogous analysis by a different group did not detect VSV-G in COPI vesicles (105). An in vitro assay reconstituting COPI vesicle budding from rat liver Golgi generated COPI vesicles that also contained the anterograde cargo polymeric IgA receptor (100). The complexity of the COPI vesicle populations remains to be explored, and the development of means to selectively isolate specific populations of COPI vesicles coupled with proteomic analyses is likely to define the cargos and the directions of distinct COPI classes.
COPI is recruited to membranes by activated ARFs. Based on amino acid sequence similarity, the six mammalian ARFs have been subdivided into three classes (79). Class I [ARF1, ARF2 (this ARF has been lost in humans), and ARF3] and class II (ARF4 and ARF5) localize to compartments of the secretory pathway and function therein. The sole class III ARF (ARF6) appears to be involved in endocytosis and actin dynamics (40).
COPI appears to be recruited to membranes by class I and II ARFs, as COPI dissociates from all cellular membranes when a GEF that activates these ARFs is inhibited by treatment with the inhibitor Brefeldin A (see below). However, it is likely that different ARFs regulate COPI recruitment at distinct compartments. This hypothesis is suggested by the slightly different localizations of the ARFs. For example, ARF1 seems more generally distributed, whereas ARF4 appears to be concentrated at the ERGIC (37). Additionally, siRNA-mediated depletion of different ARFs results in distinct effects on trafficking (184). ARFs recruit COPI coatomers by interacting directly with the β, γ, ε, β′, and δ subunits of the trimeric and the tetrameric COPI subcomplexes (49).
Like all small GTPases, ARFs cycle between a GDP-bound inactive state and a GTP-bound active state. ARF activation is mediated by a family of GEFs characterized by a conserved Sec7 domain (76). The ∼200-amino acid Sec7 domain was initially identified in the yeast protein Sec7p, and it alone is sufficient for exchange of GDP for GTP on ARF in vitro. Eukaryotic ARF GEFs are divided into five families based on overall structure and domain organization: GBF/BIG, cytohesins, EFA6, BRAGs, and F-box.
GBF1 is the sole ARF activator required for COPI recruitment to membranes and COPI vesicle formation. Inactivation of GBF1 with the specific inhibitor Golgicide (GCA) (138) or RNAi-mediated depletion of GBF1 (103, 175) causes COPI dissociation from membranes. GBF1 has been shown to activate ARF1, ARF4, and ARF5, but not ARF3, despite strong similarity to ARF1 (102). GBF1 has been localized to ERGIC and Golgi where it co-localizes with COPI. GBF1 interacts with Rab1b, and this interaction appears to promote GBF1 recruitment to membranes (110). It is possible that GBF1 activates distinct ARFs at distinct compartments to facilitate formation of vesicles coated with different coatomers and containing different cargoes.
Sorting of type I transmembrane proteins that contain NH2 termini within the ER lumen and COOH termini within the cytoplasm into COPI vesicles for recycling to the ER is mediated by two highly conserved motifs within the COOH terminus of their cytoplasmic domains: WXXW/Y/F and KKXX. These motifs are recognized by the α, β′, γ, and δ subunits of COPI subcomplexes that mediate their incorporation into nascent COPI vesicles (38).
COPI-binding motifs are absent in all known Golgi glycosyl-transferases, all of which are type II proteins with cytosolic NH2 termini. Thus glycosyl-transferases must utilize a distinct way to be incorporated into recycling COPI vesicles. Recent studies showed that the cytosolic tail of the KRE2 α-1,2-mannosyl transferase of the medial-Golgi binds Vps74p and that this interaction is required for the Golgi localization of this enzyme (148, 181). Importantly, Vps74p is not required for the Golgi localization of other Golgi proteins such as the multi-spanning Rer1p and Tvp15p. Thus Vps74 appears to be a COPI vesicle "loader" specific for only glycosyl transferases. Vps74 appears to bind the semi-conserved motif of F/L-L/V/I-X-X-RK that is found in the cytoplasmic NH2 termini of the known 16 Golgi glycosyl-transferases (181). Mutation of the FLS motif in Kre2p to AAA abolished Vps74 binding and abolished Kre2p localization to the Golgi, but it remains to be determined whether Vps74 sorts all glycosyl transferases into COPI vesicles. Vps74 links the glycosyl transferases to the COPI coat by binding to the Sec26p (β-COP) and the Ret2p (δ-COP) of the coatomer (181). A mammalian homolog of Vps74p is GMx33α and GMx33β (also called GOLPH3, GPP34, and MIDAS), shown previously to bind Golgi glycosyl transferases (160, 195).
In addition to transmembrane proteins that recycle back to the ER by either directly interacting with the COPI machinery or using the Vps74 linker, soluble proteins that contain the KDEL COOH-terminal motif also are retrieved via COPI vesicles. The KDEL sequence is recognized by the KDEL receptor, a seven-span transmembrane protein that continuously cycles between the ER and the Golgi (180). KDEL receptor directly interacts with COPI components (99). Three human homologs of KDEL receptor exist (ERD21, ERD22, and ERD23) and appear to have overlapping but distinct substrate preferences (132). Cargo binding by KDEL receptors is regulated by differences in the luminal pH within the Golgi (binding is promoted by the relatively low pH of ∼6.4) and the ER (dissociation is promoted by the higher pH of ∼7.2) (144).
The formation of COPI vesicles also appears to involve the activity of ARF GAPs (78, 89, 96, 97, 117, 118, 131, 188). ARF GAPs are characterized by a conserved GAP domain of ∼70 amino acids that includes a zinc finger motif and an invariant arginine residue. There are more than 30 genes with a predicted GAP domain in the mammalian genome. Most GAPs are large, multi-domain proteins that, in addition to inactivating ARFs, may function as scaffolds to regulate the assembly of large protein complexes. ARFGAPs are recruited to budding COPI vesicles by direct binding to active ARF, cytoplasmic tails of cargo proteins, and the coatomer (ARFGAP binds directly to the γ and β′ subunits of COPI). These interactions generate a complex consisting of cargo, coat, ARF, and ARFGAP. ARFGAP activity appears to be stimulated by cargo and coat proteins and leads to increased GTP hydrolysis by ARF and release of ARF from the complex. This coordination between cargo, coats, and ARFGAP ensures that only vesicles containing cargo proteins are produced (64, 96). It remains unclear whether ARFGAP1 remains as a component of mature COPI vesicles, but it is accepted that GTP hydrolysis by ARF is insufficient to cause coat dissociation. The mechanisms that regulate COPI vesicle uncoating remain to be characterized. The existence of multiple GAPs to regulate ARF activity and the currently accepted model that GAPs do not persist on budded COPI vesicles is quite different from the Sec23 subunit of COPII coat that functions as GAP for Sar1 and is incorporated into budded COPII vesicles.
A number of mammalian ARFGAPs have been localized to the Golgi including ARFGAP1 and the closely related ARFGAP2/3, ASAP2, ARAP1, and SMAP2. It remains to be determined whether distinct ARFGAPs regulate the formation of distinct COPI vesicles containing different sets of proteins.
Defects in COPI function cause disease or developmental abnormalities, highlighting the essential role of this coating machinery. A single amino acid substitution of Ile 422 to Thr in the δ COP subunit close to the cargo-binding site causes coat-color dilution and ataxia due to degeneration of Purkinje cells in the cerebellum in mice (197). The mutation disrupts intracellular trafficking in melanocytes, but the exact cargoes that are mistargeted in the melanocytes and the brain remain to be identified. More importantly, the phenotypic severity varies in different tissues, suggesting that the trafficking of some cargoes is more affected or that some cargoes are more important to the animal's well being.
Similar differences in phenotypic severity are observed in Drosophila with loss-of-function mutations in γ COP (65). Although γ COP activity is essential for embryo viability, the most pronounced defects are observed in the morphogenesis of the epidermis and tracheal tubes. The tracheal defect correlates with defects in the secretion of the protein Piopio. This again stresses that phenotypic effects of removing components of general traffic machinery are surprisingly specific.
The function of ζ COP isoforms has been examined indirectly in a study showing transcriptional silencing of the ζ2 isoform in a number of cancer cell lines of diverse origin (158). Interestingly, this makes the tumor cells exquisitely dependent on ζ1 function. Although normal cells show normal growth after depletion of either ζ1 or ζ2 (but not when both isoforms are depleted), tumor cells are killed when only ζ1 is depleted because they do not have ζ2. This suggests that ζ1 and ζ2 can compensate for each other's function in cells in culture, although the effects of inactivating ζ1 or ζ2 in animals remain to be determined. Importantly, the strict dependence on ζ1 in cancer cells raises the exciting opportunity for developing anti-cancer therapeutics based on the solved structure of ζ COP (202).
Additional regulators of COPI that cause disease also have been identified. Scy1-like (Scyl1) is a member of the Scy-like family of catalytically inactive protein kinases and has been shown to bind β COP (28). The binding of the cytosolic Scyl1 to β COP is through a COOH-terminal RKLD sequence, similar to the KKXX motif present in transmembrane proteins recognized by the COPI machinery. Scyl1 co-localizes with COPI in cells, and depletion of Scyl1 causes defects in trafficking of the KDEL receptor. Mice with null mutations in the Scyl1 gene exhibit neuromuscular atrophy and spinocerebellar ataxia (146).
The Drosophila homolog of GBF1 called Garz is essential for embryonic viability (FlyBase). Tissue-specific silencing of Garz within the salivary gland causes a severe disruption in gland development and alteration in polarity due to arrest in surface deployment of cell-cell and cell-matrix proteins required for morphogenic events (174).
Tethering and Fusion of COPII and COPI Vesicles
The final stages of cargo delivery to the target membrane involve at least two events: recognition between the membranes of the cargo carrier and the target compartment, and overcoming the thermodynamic barrier to membrane fusion. The initial carrier-membrane interaction appears to be mediated by tethers, whereas the subsequent fusion of the bilayers is facilitated by SNAREs.
A large family of tethering factors has been characterized (173, 192). Tethers are very diverse but can be divided into a group of coiled-coil proteins (known as golgins) (130) and a group of multi-subunit complexes (120). The coiled-coil tethers p115 (yeast Uso1p), GM130 (yeast Bug1), giantin, golgin-84, and CASP, and the multi-subunit complexes Dsl1, TRAPPI, TRAPII, and COG are all involved in ER-Golgi trafficking.
Tethering COPII vesicles.
Initial insight into the tethering mechanisms for COPII vesicles came from yeast. The yeast Uso1p was shown to tether COPII vesicles to yeast Golgi in an in vitro reconstituted system, and Uso1p is required for COPII vesicle tethering in vivo (10, 33, 34). Uso1p and its mammalian homolog p115 (described below) are large peripherally membrane-associated proteins that exist in vivo as parallel homodimers with two globular heads and a long coiled-coil tail (115, 151, 199). Interestingly, the tails consist of four coiled-coil motifs separated by proline-rich "hinges" that have been proposed to allow the accordion-style collapse of the tethers, perhaps to bring the vesicle into close proximity to the target membrane.
Recruitment of Uso1p to COPII vesicles appears to be regulated by the active form of the small GTPase Ypt1p (33). Uso1p presumably links the COPII vesicle to Golgi membrane by binding to a Golgi protein, but the exact binding partner on the Golgi is uncertain. Defects in Uso1 can be suppressed by overexpression of Ypt1 and the known ER-to-Golgi SNAREs (Bet1, Bos1, Sec22, and Ykt6), and the mammalian p115 binds SNAREs, raising the possibility that Uso1p might link COPII vesicles to Golgi membranes by binding to a Golgi SNARE.
In addition to Uso1p, other proteins participate in COPII vesicle tethering. Recent studies indicate that yeast Grh1p, a relative of GRASP65 (described below) also associates with COPII vesicles by binding the Sec23/Sec24 subunits of the COPII coat (15). Grh1p appears to facilitate COPII vesicle tethering because Grh1p depletion inhibits tethering in an in vitro reconstituted assay (15). The exact mechanism by which Grh1p functions is unknown, but Grh1p binds the coiled-coil protein Bug1p (which is structurally similar to GM130, a known GRASP65 interactor), and it is possible that this interaction is involved in tethering. Bug1p is also required for COPII vesicle tethering in vitro (15). As is the case for Uso1p, the identity of the binding partner on the Golgi for Grh1p and Bug1p is unknown. It also remains to be determined whether Uso1p, Grh1p, and Bug1p function together in a common pathway or in separate pathways to facilitate COPII vesicle tethering. Based on findings from the mammalian system (see below), it is likely that they act together.
The mechanism for tethering COPII vesicles in mammalian cells is likely to involve p115, GM130, and GRASP65. However, the target of COPII vesicles in mammalian cells is less clear, and it is possible that COPII vesicles tether to pre-Golgi compartments, possibly ERGIC and maybe also the Golgi (3, 112, 196).
p115 localizes to COPII vesicles, and p115 recruitment to COPII vesicles is promoted by the active form of the mammalian Ypt1 homolog Rab1 (3). p115 also localizes to ERGIC and the Golgi, and this association is extremely dynamic (25). p115 binds the Golgi-localized coiled-coil tether GM130 (92), and it is possible that this interaction facilitates the tethering of COPII vesicles to Golgi membranes. GM130 is also present on ERGIC (104) and could mediate COPII vesicle tethering to that compartment. GM130 is a peripherally associated protein that is bound to Golgi membranes by the Golgi reassembly stacking protein of 65 kDa (GRASP65) (12). GRASP65 is N-myristoylated, and this mediates the tight association of GRASP65 with membranes (12). Thus p115 on incoming COPII vesicles may bind to GM130-GRASP65 tightly associated with ERGIC and Golgi membranes to tether COPII vesicles.
In addition to coiled-coil tethers, COPII vesicle tethering in yeast also involves the heptameric TRAPPI complex (30). TRAPPI is unique because it acts as a Rab GEF (32). Yeast TRAPPI activates Ypt1, whereas mammalian TRAPPI activates Rab1 (186). Yeast TRAPPI appears to tether ER-derived COPII vesicles through a direct interaction between the Bet-3 subunit of TRAPPI and the Sec23 subunit of COPII (30). The participation of the coiled-coil tethers and the heptameric TRAPPI appears to be sequential: TRAPPI binds to Sec23p on budded COPII vesicles and activates Ypt1/Rab1 on the vesicles, and this recruits Uso1p/p115 to the vesicles (30). Uso1/p115 then tethers COPII vesicles to Golgi, presumably by interacting with Bug1/Grh1/GM130/GRASP65.
Tethering COPI vesicles.
Tethering COPI vesicles is significantly more complex because, unlike COPII vesicles that originate from a single compartment (the ER) and deliver cargo to a single compartment (the Golgi), COPI vesicles bud from many compartments and fuse with many compartments. Tethering of COPI vesicles to the ER appears to be mediated by the trimeric Dsl1 complex composed of the soluble proteins Dsl1, Dsl3, and Tip20 (107). Dsl1 localizes to the ER by stably associating with the ER SNAREs Ufe1, Use1, and Sec20 (107). Dsl1 has been shown to interact with the α and ε subunits of the trimeric subcomplex of the coatomer that forms the outer layer of the COPI coat (4, 5, 73, 133, 182). Thus interaction between Dsl1 and the outer layer of the COPI coat is optimal for "catching" recycling COPI vesicles and facilitating their fusion with the ER.
Intra-Golgi COPI vesicles appear to be tethered by multiple tethers, depending on their destination. The p115 tether has been proposed to mediate COPI vesicle tethering to cis- and perhaps medial-Golgi because p115 is restricted in its localization to those Golgi compartments (116). In support of the role at that level of the Golgi, p115 binds the cis-Golgi protein GM130 (stably associated with Golgi membranes through GRASP65) and the medial-Golgi transmembrane coiled-coil protein giantin (91, 163). p115 binds the β-COP subunit of COPI (66) and thus could directly tether COPI vesicles derived from medial-Golgi and to cis-localized GM130 and COPI vesicles derived from trans-Golgi to medial-localized giantin.
Golgin-84 is a transmembrane coiled-coil protein localized to medial-Golgi (13, 46). Overexpression or depletion of golgin-84 causes fragmentation of the Golgi (46) and causes a partial mislocalization of Golgi proteins into adjacent cisterna and accumulation of untethered COPI vesicles (161). However, injection of soluble golgin-84 into cells does not affect the retrograde movement of ERGIC53 from the cis-Golgi to the ER (100). Together, the data suggest that golgin-84 mediates tethering exclusively within the Golgi. Golgin-84 binds CASP, a coiled-coil protein localized to medial-Golgi (60). COPI vesicles generated in an in vitro budding assay contain golgin-84 and can be recovered on CASP-coated glass beads (100), strongly suggesting a tethering function.
Both p115 and golgin-84 appear to intersect with the octameric COG tethering complex (161, 162). COG localizes to the Golgi where it mediates the tethering of COPI vesicles that traffic in the retrograde direction between Golgi cisterna (153, 159, 209). Depletion of Cog3 (and the corresponding decrease in the octameric COG complex) leads to the accumulation of nontethered COPI vesicles containing Golgi SNAREs and Golgi resident proteins (209). COG interacts with p115 (162), COPI subunits (172, 209), and SNAREs (153, 172) and may directly participate in stabilizing SNARE complexes (153), a function it shares with other tethers (see below). Golgin-84 interacts with the Cog7 subunit of COG, and preventing this interaction by expressing the Cog7-interacting domain of golgin-84 disrupts Golgi morphology in a manner consistent with inhibition in intra-Golgi traffic (161). However, the exact origins and destination of COPI vesicles tethered by COG and how COG function coordinates with p115 and golgin-84 (and by extrapolation with GM130, GRASP65, giantin, and CASP) remain under investigation.
Mutations in subunits of COG have been shown to cause often-fatal congenital disorders of glycosylation type II (CDG-II) in humans. A defining defect in CGD-II is improper glycosylation of N- and O-linked glycoproteins. An 80-amino acid truncation of the COOH terminus of Cog1 causes reduced levels of cellular COG complex and results in mild CDG-II (55). Another mild form of CDG-II was detected in a patient with an R729W substitution in Cog4 that caused reduced levels of the assembled octameric COG complex (135). A moderate CGD-II characterized by psychomotor retardation was observed in a patient with reduced expression of Cog5 (125). In contrast, a G549V substitution in Cog6 that significantly decreased the levels of octameric COG complex resulted in a neurological disease with a fatal outcome in early infancy (94). Similarly severe phenotypes were observed in patients with a mutation in Cog7 that significantly decreased overall COG levels (111, 203). A 76-amino acid truncation of the COOH-terminus of Cog8 that prevents the assembly of heptameric COG complex but allows the assembly of COG subcomplexes caused a milder neurological phenotype (54, 83). Overall, it is obvious that even a small decrease in cellular levels of the fully assembled octameric COG complex are deleterious.
In addition to COG, another multi-subunit complex, TRAPPII, has been suggested to facilitate COPI vesicle tethering within the Golgi. Yeast TRAPPII appears to be composed of nine subunits, six of which are shared with TRAPPI and three that are TRAPPII-specific (36, 200). In yeast, the role of TRAPPII complex appears to be largely at the TGN and within the endosomal system (31). A single mammalian TRAPPII has been described and appears similar to the TRAPPII complex from yeast in that it also contains the TRAPPII-specific Trs130, Trs120, and Trs65 subunits (137). Mammalian TRAPPII localizes to the ERGIC and the Golgi, and within the Golgi appears enriched on COPI-coated buds and vesicles but not on cisternal elements. Depletion of Trs130 disrupts Golgi morphology and leads to the accumulation of vesicles adjacent to the Golgi. It also inhibits trafficking of VSV-G at the disrupted Golgi, suggesting that TRAPPII function is required within the Golgi after ER to Golgi step of transport. Importantly, the Trs130 and the Trs120 subunits of TRAPPII bind γ1 COP. This suggests that TRAPPII could directly link COPI vesicles to target membranes. However, TRAPPII has GEF activity for Rab1, and it is possible that TRAPPII functions within the Golgi to activate Rab1 (to allow recruitment of Rab1-dependent tethers such as p115), and is not a bona fide tether.
All tethers are either long proteins or large assemblies that appear perfect for "catching" vesicles from a long distance. Coiled-coil tethering factors have been proposed to act by assembling tethering bridges that originate from vesicles and from acceptor membranes, whereas multi-subunit tethering factors localize to and act predominantly on the acceptor membranes (98). The model of tethers as molecular bridges has been refined, and it is possible that tethers interact with other components of the fusion machinery to directly facilitate fusion. Specifically, tethers have been shown to directly bind SNAREs and to facilitate the formation of SNARE pins as a prelude to fusion (45, 153, 156). Thus tethers are believed to accelerate a rate-limiting step in SNARE complex assembly. Tethers also bind Rabs, and this interaction may increase tethering specificity or position the Rab in close proximity to the SNAREs and promote their pairing. The exact role of Rabs in fusion is still debated, but the requirement for Rab function is absolute (29, 88). Tethers might represent molecular platforms that coordinate the function of Rabs and SNAREs (and perhaps other factors) in fusion.
Membrane-membrane fusion during vesicular traffic is facilitated by members of a large family of SNAREs (27, 52, 72). Members of the SNARE family are found on vesicles (v-SNAREs) and on target membranes (t-SNAREs), and fusion of the two opposing membranes occurs through pairing of cognate SNAREs. Assembly of SNARE complexes involves the formation of a 4-α-helix bundle, where one helix is contributed by the v-SNARE and three by t-SNAREs (53, 190). The interaction of v- and t-SNAREs on the vesicle and target membrane is described as the trans-configuration or trans-SNARE. This configuration brings the two membranes into sufficient proximity for membrane fusion. After membrane fusion, the v- and t-SNAREs are still together but now reside within the same membrane, a conformation referred to as cis or cis-SNARE. The cis-SNARE complexes are disassembled in an ATP-dependent process through the action of the N-ethylmaleimide-sensitive factor (NSF) (113, 114, 124, 191, 201). This disassembly step regenerates the cellular pool of free SNAREs and allows repeated rounds of transport.
It is extremely difficult to define the membranes in which individual SNAREs function in ER-Golgi traffic because these proteins continuously cycle between compartments (39, 69, 77, 198, 207). A complex of syntaxin-5, Sec22b, membrin, and Bet1 appears to facilitate membrane fusion at early steps, probably catalyzing fusion of COPII vesicles with the ERGIC. A distinct complex composed of syntaxin-5, GOS28, Bet1, and Ykt6 acts later, perhaps to facilitate fusion of anterograde carriers at the cis-Golgi. A third complex composed of syntaxin-5, GOS28, Ykt6, and GS15 acts late within the Golgi, perhaps facilitating intra-cisternal traffic.
Future Directions
Despite intense interest and experimentally derived insights, many questions remain about protein trafficking and activity of proteins involved in this process. Some of the more intriguing points are listed below.
1) It remains unknown how many different types of COPII vesicles bud from the ER and whether different cargoes are packaged into different COPII vesicles. Bioinformatics of mammalian proteomes suggest that of the ∼25,000 predicted open reading frames, ∼11% (2,750) encode soluble secretory proteins and ∼21% (5,250) encode transmembrane proteins destined for compartments of the secretory and endosomal systems (35, 81). Although not all genes are expressed in all cells, it is reasonable to predict that a typical cell must efficiently export thousands of proteins out of its ER by packaging them into COPII vesicles. How different cargos are handled for export in different cell types remains to be elucidated.
2) It is also unknown how many distinct types of COPI vesicles are generated from the Golgi and the ERGIC, and how all these vesicles are correctly targeted to the compartment that is proximal to their compartment of origin.
3) The increased complexity of the mammalian secretory pathway is paralleled by the evolution of isoforms for most of the key players, and there are multiple isoforms of the GEFs, GAPs, Sar/ARFs, and COPII and COPI subunits. However, how these different isoforms function in different tissues, different stages of traffic, and trafficking of select cargos remain to be determined.
4) The timing of coat disassembly and the in vivo triggers for uncoating remain to be characterized. Interestingly, the interactions between COPII and COPI coats and tethers (p115, Dsl1, COG, and TRAPPI) suggest that tethering occurs before vesicles uncoat. Furthermore, Dsl1 has been shown to bind to sites involved in interactions between COPI subunits that stabilize the COPI coat (134, 147, 208). Thus Dsl1 might initiate the uncoating of COPI vesicles tethered to the ER. This raises the question of whether other tethers also may aid in uncoating, how coats are shed while bound to tethers, and how the uncoated vesicles are prevented from escaping.
5) Finally, one of the key questions in membrane trafficking involves upstream regulatory mechanisms. The secretory pathway responds to organismal demands for secretion during events such as development, immune challenge, lactation, etc. The signaling and transcriptional mechanisms that facilitate amplification of the secretory pathway are under active investigation, and analysis of early acting transcription factors required for the development of the salivary glands (an organ with extreme secretory capacity) in Drosophila uncovered a key role for a complex of homeotic transcription factors Scr/Exd/Hth (Sex Comb Reduced/Extradenticle/Homothorax) (1). This complex directly regulates the CrebA transcription factor that alone is sufficient and necessary for upregulation of at least 34 genes involved in the biogenesis of the secretory pathway and secretion (56). CrebA not only induces expression of protein machinery involved in secretion at the ER-Golgi interface but also more than 20 known or predicted cargo proteins. Microarray analysis of HeLa cells expressing Creb3L1 (Creb3L1 and Creb3L2 are the mammalian orthologs of Drosophila CrebA) shows upregulated expression of genes involved in Golgi vesicle transport and secretion (56). Thus Creb3L1 can turn a HeLa cell that is not specialized for secretion into a cell that amplifies the secretory pathway. The important role of the CrebA-/Creb3-like transcription factors in regulating the secretory pathway is also underscored by the recent finding that a missense mutation in Creb3l2 (a zebra fish ortholog of CrebA/Creb3) decreases the expression of Sec23a, Sec23b, and Sec24d subunits of the COPII coat (108).
Recent advances have revealed significant insight into the complexity of membrane trafficking. Continuing molecular, structural, biophysical, and reconstitution studies in vitro and in vivo will further our understanding of how the trafficking machinery functions to facilitate efficient and high-fidelity transport of cargo through multiple secretory compartments.
We thank Drs. Guillermo Marques, Rafael Garcia-Mata, Cecilia Alvarez, and Melanie Styers for constructive comments on the manuscript.
This work was supported in part by grants from the National Institute of Health, the National Science Foundation, and the American Heart Association.
No conflicts of interest, financial or otherwise, are declared by the author(s).
- ©2011 Int. Union Physiol. Sci./Am. Physiol. Soc.
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text:
Dormy Inn Himeji
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Dormy Inn Himeji
About the Hotel
Property Location
Located in Himeji, Dormy Inn Himeji is minutes from Kasuga Shrine and close to Odoshi Shrine. This hotel is within close proximity of Shimmei Shrine and Junisho Shrine.
Make yourself at home in one of the 156 air-conditioned rooms featuring refrigerators. Complimentary high-speed (wired) Internet access keeps you connected, and satellite programming is available for your entertainment. Bathrooms have complimentary toiletries and hair dryers. Conveniences include phones and coffee/tea makers, and you can also request irons/ironing boards.
Rec, Spa, Premium Amenities
Make use of convenient amenities, which include complimentary wireless Internet access and tour/ticket assistance.
You can enjoy a meal at a restaurant serving the guests of Dormy Inn Himeji, or stop in at a grocery/convenience store.
Business, Other Amenities
Featured amenities include complimentary high-speed (wired) Internet access, a computer station, and dry cleaning/laundry services. Self parking (subject to charges) is available onsite.
Hotel Facilities
- Laundry facilities
- Tours/ticket assistance
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- Total number of rooms - 156
- Computer station
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- Free WiFi
- Year Built 2011
- Parking (limited spaces)
- Breakfast available (surcharge)
- Free wired high-speed Internet
- Grocery/convenience store
Room Facilities
- Satellite TV service
- Air conditioning
- Slippers
- Hair dryer
- Coffee/tea maker
- Television
- Free toiletries
- Pay movies
- Phone
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- Free wired high-speed Internet
Hotel Policy
Know Before You Go
- The Japanese Ministry of Health, Labour, and Welfare requires all international visitors to submit their passport number and nationality when registering at any lodging facility (inns, hotels, motels, etc. ). Additionally, lodging proprietors are required to photocopy passports for all registering guests and keep the photocopy on file.
The following fees and deposits are charged by the property at time of service, check-in, or check-out.
- Self parking fee: JPY 1000 per day
The above list may not be comprehensive. Fees and deposits may not include tax and are subject to change.Show all hotel policy Show less hotel policy


text:
|Hardcopy (shipping and handling)
Work Item(s) - proposed revisions of this standard
More F13.30 Standards Related Products Standard References Reprints and Permissions
Significance and Use
This non-proprietary laboratory test method allows preliminarily for the reproducible testing of whole footwear and footwear-related soling materials for evaluating relative slip performance. Other ASTM test methods generally employ a standardized test foot primarily for evaluation of flooring materials.
FIG. 1 Example of Footwear Mounted Using 7° Wedge to Set Proper Contact Angle
1. Scope
1.1 This test method determines the dynamic coefficient of friction between footwear and floorings under reproducible laboratory conditions for evaluating relative slip performance. The method is applicable to all types of footwear, outsole units, heel top-pieces (top-lifts) and sheet soling materials, also to most types of indoor floorings, including matting and stair nosing, and surface contaminants on the flooring surface, including but not limited to liquid water, ice, oil and grease. The method may also be applied to surfaces such as block pavers, turf and gravel.
1.2 Special purpose footwear or fittings containing spikes, metal studs or similar may be tested on appropriate surfaces but the method does not fully take account of the risk of tripping due to footwear/ground interlock.
1.3 The values stated in the ASTM test method in metrics are to be regarded as the standard. The values in parentheses are for information.
1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.
2. Referenced Documents (purchase separately) The documents listed below are referenced within the subject standard but are not provided as part of the standard.
ASTM Standards
F1646 Terminology Relating to Safety and Traction for Footwear
Other ReferencesEN 10088-2 Stainless steels - Part 2: Technical delivery conditions for sheet/plate and strip of corrosion resisting steels for general purposes Available from European Committee for Standardization (CEN), Avenue Marnix 17, B-1000, Brussels, Belgium, http://www.cen.eu.
ICS Code
ICS Number Code 61.060 (Footwear); 97.150 (Non-textile floor coverings)
UNSPSC Code
UNSPSC Code 46181600(Safety footwear)
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Citation Format
ASTM F2913-11, Standard Test Method for Measuring the Coefficient of Friction for Evaluation of Slip Performance of Footwear and Test Surfaces/Flooring Using a Whole Shoe Tester, ASTM International, West Conshohocken, PA, 2011, www.astm.orgBack to Top


text:
- Description
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- Suitable for (7)
- Reviews
RMS, Italy, is a prolific manufacturer of OEM quality aftermarket parts for many scooter brands, including Vespa Vintage and Vespa Modern parts. Their catalogue is extensive, with equipment made for even the most vintage of models.
This is an RMS Clutch Push Rod Kit for Vespa PE 200cc. This kit including pressure plate, circlip and push rod will replace the worn down clutch parts. Used and worn down clutch components lead to loss of clutch grip and results in poor overall performance.
For VSX - VSX2
Data Sheet
Name RMS Clutch Repair / Maintenance Kit Vespa PE 200cc (VSX - VSX2) Item No. R100300171 Brand RMS
Your product goes with the following manufacturers and their models:
Vespa Classic
PE 200cc 1977 - 1984 (VSX1T)
PX Cosa 200cc P E 1980 (VSX1T)
PX Cosa 200cc PX E 1981-1983 (VSX1T)
PX Cosa 200cc PX E LUSSO 1984 - 1997 (VSX1T)
PXE 200cc 1977 - 1984 (VSX1T)
PXE 200cc Arcobaleno 1983 - 1984 (VSX1T)
PXE 200cc Arcobaleno Elestart 1984 (VSX1T)


text: Arstechnica.com How I learned to stop worrying and love the big $60B NASA rocket The Space Launch System's complete qualification hydrogen fuel tank, ready to be shipped by barge to Marshall Space Flight Center for testing. Eric Berger Pressure domes for SLS hydrogen tanks. Eric Berger Michoud Assembly Facility goes on and on. It encompasses 43 acres under a single roof. This is where NASA is building the Intertank, the connection... #space #nasa #eric-berger Tweet Source: arstechnica.com Read More


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SAN DIEGO-- Jeff Moorad is set to become the Padres' nextowner.
In a deal that was confirmed Tuesday and had been rumored for weeks, Moorad's ownership group is expected to purchase full ownership of the team from current owner John Moores within five years. A source with knowledge of the negotiations who is not authorized to speak publicly about the sale said Moorad's group is currently purchasing one-third of the team and the deal could approach $550 million.
The agreement coincides with Moores' divorce from his wife,Becky, and still is pending the approval of Major League Baseball.But Moorad, who resigned as the Arizona Diamondbacks' CEO lastmonth to pursue the purchase of the Padres, presumably will takeover as the club's CEO by Opening Day 2009.
Moorad's arrival also will signal the departure of current CEOSandy Alderson, who confirmed Tuesday he will resign when Mooradtakes control. Moorad -- who wouldn't reveal the names of hispartners or the amount they are contributing to the purchasebecause not all have passed through MLB's approval process -- alsosaid Moores will remain the team's "control person" and actingchairman.
"It's an exciting day and a large step forward in our process of acquiring the Padres," Moorad said. "We have one more hurdle to go, but we're looking forward to completing this transaction in the next couple of months."
The deal, which includes ownership of the team and its 30 percent stake in Petco Park, could be completed in as early as one year. However, Moorad said his group likely would use all five years agreed to by Moores to complete the purchase. Last April, Forbes magazine estimated the club's value at $385 million. Moores purchased the team from Tom Werner's group in 1994 for $80 million.
"Jeff Moorad will not only bring to the Padres his unique experience, passion for the game, and proven leadership, but an unparalleled commitment to the City of San Diego and its dedicated fans," Moores said in a statement. "I speak for our entire organization in saying that we look forward to working with Jeff to reach the ultimate goal of winning a World Series title for America's finest city."
The Padres are coming off a 99-loss season, their worst in 15years. Under Moores' direction, the team also has spent the pastseveral weeks shedding salary, with the payroll expected to drop to$40 million for 2009. Most notably, the team declined to re-signlongtime closer Trevor Hoffman. The Padres also have been trying totrade pitcher Jake Peavy, their best player. Peavy is owed $63million over the next four seasons.
A former sports agent of more than 20 years whose clients included Steve Young, Troy Aikman and Manny Ramirez, Moorad was a minority partner in a group that purchased the Arizona Diamondbacks in 2005. Taking over as Arizona's CEO at the time, many baseball insiders, from owners to players' union representatives, feared that a former agent would manipulate the system by using insider information he gained during his years as a player representative. But Moorad has done much to overturn that skepticism, including leading the Diamondbacks out of their serious financial troubles. When Moorad took over, the team was $353 million in debt. Now, however, the club turns a small profit and is paying down its debts while remaining competitive on the field.
Many baseball insiders see Arizona and San Diego as similar markets because of their size. While running the Diamondbacks, Moorad preferred to build the organization through development of a strong farm system while shying away from spending big money on the free agent market. During Tuesday's conference call, Moorad suggested that under his tenure the Padres would operate in a similar fashion.
"I think the model will be similar," Moorad said. "They're certainly will be similarity between the approaches. The revenue is the same and the state of the Padres is similar to the Diamondbacks a couple years back. But I want to impress upon the fans, we do have a plan. … It will be a plan that requires patience on the one hand but designs that the team will have a competitive ballclub on a consistent basis."
Contact staff writer Dan Hayes at email@example.com .


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Jerusalem, Oct 9 (IANS) Israeli and Russian military officials have met in Tel Aviv to sort out a mechanism that would prevent accidents between their forces as Russia continues its operations in Syria, the Israel Defence Forces (IDF) has said.
During the past week, meetings were held between Russian and Israeli delegations, headed by the deputies of chief of staffs of both countries, the IDF said in a statement released on Thursday night, Xinhua reported.
"The meetings dealt with establishing a mechanism of safety between both militaries in the region," IDF said, with a follow-up meeting scheduled to take place in Moscow in the upcoming weeks.
Russian Deputy Chief of Staff Nikolai Bogdanovski arrived in Tel Aviv on Tuesday with a delegation of military officials. His Israeli counterpart, Yair Golan, hosted the meetings, two weeks after Israeli Prime Minister Benjamin Netanyahu and Russian President Vladimir Putin agreed on the coordination during a meeting in Moscow.
Prior to the meetings, the IDF stated that the talks would focus on coordinating the activity of both armies in the "aerial, naval and electro-magnetic arenas."
An Israeli defence official told the Walla! News website the initial meetings consisted of introductions and establishing a common ground, before actually starting to sort out its details.
He said that, apart from the risk of incidental clashes between the forces, there is also concern in the military that the Russian operation in the Syria would disrupt the operation of Israel's radars deployed along the Syrian border.
Russia has been conducting air strikes in the past week in Syria against the Islamic State (IS). It boosted military presence in Syria which has been mired in a bloody civil war since 2011, claiming that it is aimed at crushing IS militants.
However, the US and its allies are concerned that Russia's actions are not only aimed at defeating IS, but also against militant groups supported by the U.S. in their battle against the Syrian government.
Israeli officials have repeatedly stated that Israel does not intervene in the ongoing civil war in Syria, but rather act to defend Israel's interests.