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Not since FDR has america been confronted with the enormity of rebuilding America's financial system and infrastructure. In the present day our financial system in addition to the nations infrastructure is placing this nations safety and stability in grave hazard. Our financial system at this time is already on the verge of falling into the abyss by elements very paying homage to the occasions that led to the Nice Melancholy of the 1930's. And, sure one other housing bubble just like the one we simply had in 2008 is poised to burst. However, you wouldn't understand it the way in which Trump is citing his most illustrious accomplishments and naturally the media is correct behind with their bogus reporting of the unemployment numbers. What we're confronted with at this time is that america remains to be woefully unprepared to deal with any monetary calamity.To high it off our "Wizards" on capital hill and Trump within the White Home proceed to be oblivious to the calamity of both a infrastructure catastrophe or the continued financial woes which might be protecting thousands and thousands of American's languishing in monetary devastation would do to this nation.
All prepared in virtually each metropolis throughout the nation there's a Division Of Financial Growth. The first focus of this division is to generate extra enterprise development inside their respective metropolis. In doing so extra tax income would stream into these cities to assist the mandated public companies that are actually required by legislation. With out enterprise development as in so many situations at this time all throughout the nation metropolis finances shortfalls solely exasperate an ongoing financial domino impact of monetary hardship for all. There are numerous elements that must be applied to safe enterprise development to extend and increase the tax base inside every metropolis all throughout the nation. With out extra instructional venues provided, accessible dependable public transportation out there, reasonably priced first rate housing and all the opposite logistics that make it not solely conducive however productive for enterprise growth in cities and cities enterprise development will proceed to say no. The tax base might be placed on the burden of the house proprietor which solely creates extra financial hardships for all.
When the Division Of Commerce and Labor was created again 1903 and subsequently consolidated in 1913 into the Division of Commerce the one time that that company has managed to spur actual financial development was via the intervention of FDR. The quite a few public works initiatives that FDR applied facilitated Americas victory in World Battle II and propelled america to it's best financial growth following the Battle via to the top of the 1960's. What has occurred since are a variety of coverage adjustments which have undermined the financial and monetary alternatives of so many thousands and thousands of People. Consequently, now we have inadvertently created the best wealth disparity hole in historical past, misplaced virtually all of our manufacturing capabilities, and diminished the center class to virtually extinction. All this with the truth that our academic ranges have plummeted in order that america isn't even on the radar kind of talking in international instructional requirements. In essence our graduates are usually are not totally ready for no matter jobs which might be out there at this time.
Now, when the financial calamity from 2008 monetary disaster hit the fallout remains to be protecting the us financial system stagnant at finest. Just some years in the past the Hostess Company closed so many bakeries they laid off over 18,000 workers simply in time for the vacations. In Florida of that very same yr over 2,000 state employees suffered the identical destiny. In the present day, the retail business is dying forcing extra folks onto the unemployment traces. With Trump within the White Home and our Republican led Congress have executed really nothing to spark a rejuvenation in employment numbers. The media nonetheless fails to inform the reality concerning the unhappy incontrovertible fact that unemployment and underemployment have decimated our nations skill to prosper. That is one more reason why there may be a lot agitation particularly inside the Black neighborhood. In each metropolis and city throughout the nation the African American unemployment fee is greater than triple that of what the media is reporting.
The fact has but to hit dwelling to our elected officers to the true disaster within the nation. The dearth of alternative for thousands and thousands of People to have the ability to earn a dwelling wage is staggering. And, but too many officers actually have turned a blind eye to what's actually happening. When now we have riots within the streets as an alternative of employees in factories, or behind counters, or in so many different occupations is a inform story signal that governmental insurance policies for the previous 40 years have undermined the American employees skill to prosper. When now we have CEO's raking in large exorbitant salaries and company earnings at file ranges whereas the remainder of us languish with hunger wages or worse but having to depend on meager unemployment advantages that don't final creates a tempest that's solely exasperated by our elected officers failure to implement insurance policies that might propel america into brighter and safer future. What now we have at this time is that our Republican led Governors, members of Congress, in addition to our notorious President can't fairly grasp what they've executed and proceed to do by not ushering insurance policies that truly put extra individuals again to work with dwelling wages very similar to what FDR did.
From all indications for the reason that election it's fairly obvious that these in Washington nonetheless, like so many Republicans, can't grasp the truth dealing with thousands and thousands of People at this time. The Division of Commerce has executed some semblance of making an attempt to reassemble financial development however this Administration nonetheless doesn't have a definitive plan of course each quick and long run that might successfully curtail the occasions like those that unfolded when the Hostess Company closed it's doorways. It's time to restructure the Division of Commerce and re institute an expanded Financial Growth Administration in order that the Division of Commerce would now turn out to be the Division Of Financial Growth and incorporate all important posts that might felicitate financial and monetary development throughout the nation.
Presently there are too many duplications in virtually each cupboard submit the place a lot wanted assets are actually being diverted from reaching the areas that want it most. It's fairly obvious that steps must be taken to reform and consolidate particularly the present Division of Commerce in order that the assets really will go the place they may do essentially the most good. This consists of putting the emphasis on Financial Growth with a significant emphasis on equalizing our commerce deficit, investing in infrastructure restoration, securing our power grid, concentrate on lowering the unemployment numbers particularly in African American communities and establishing dwelling wage requirements all throughout the nation. Not simply elevating the federal minimal wage both.
When most consider our commerce deficit many query why China is cornering the market? In virtually each shelf in virtually each retailer throughout the nation there are such a lot of gadgets that actually are made in China. However, that doesn't present the larger image. When in actuality it isn't all these imports. Positive, they contribute a lot to our lack of producing jobs however what else is curbing our financial development is correct right here in america? Training for one factor.In the present day's instructional ranges hare a far cry from what the US had again within the 1950 via the 1960's. The requirements in training have made highschool graduates not with the ability to compete for the roles which might be out there at this time. Training of our youth and younger adults is significant in turning the tide towards enhancing the unemployment numbers.
As a way to reply why China is gaining economically now we have to take a detailed have a look at our imports which have for the previous 40 years have been far exceeding our exports. To not point out what NAFTA continues to do with reference to our lack of general productiveness. We've got to keep in mind that proper after NAFTA was signed in 1993 our job losses have been staggering and the impact remains to be being felt at this time. In the present day, our complete financial state of affairs is nearly on the identical parallel because the 2008 housing disaster. The pervasive angle amongst too many CEO's overlook what Henry Ford did so many a long time in the past. He strengthened the center class whereas at this time's CEO's have solely weakened the center class to the purpose of virtually no return. A latest report reveals that wages have continued to stagnate. And adjusted for inflation at this time wages have shrunk drastically. The unhappy incontrovertible fact that at this time thousands and thousands which might be nonetheless working have seen their incomes lower whereas all the things else the associated fee maintain growing. This retardation of wages all throughout the nation is the unhappy actuality of at this time. We've got to keep in mind that the well being and stability of any nation lies with nearly all of folks incomes dwelling wages. The extra folks with sufficient disposable revenue to spend, pay down debt, and to avoid wasting a minimum of 10% yearly is the best financial increase for any nation. In essence the success of The Williams Concept of Financial Evolution. That isn't the case at this time of what's taking place in America.
What the Division of Financial Growth has to do is handle our commerce agreements and reinforce not on free commerce however equal commerce together with placing increased tariffs on items coming in from China, Mexico, Canada and even Germany to call just a few. This together with coordinating with different cupboard departments on a unified plan of Course utilizing Nationwide Financial Reform because the information to implement adjustments, ideas, and insurance policies that can in truth create the setting that's conducive for extra enterprise development. This together with creating financial alternatives that needs to be out there to each single American. One other unhappy actuality of at this time is the very fact hundreds of People have already left the nation for different international locations simply because the financial alternatives are extra plentiful exterior america. When folks discover monetary and financial roadblocks proper right here in America the place the land of alternative has shut it's doorways in so many faces is a travesty that needs to be instantly rectified.
The job at hand for the Division Of Financial Growth is to increase the tax base all throughout the nation. Work with all different businesses and Departments primarily the Division of Training to retool our academic requirements to equip our youth and younger adults with the information and the instruments obligatory in order that they'll turn out to be employable within the jobs of the 21st century. This Division is remitted to rebuild our infrastructure, roads, bridges, faculties, and different buildings that poise a risk to the protection of all. To work with the Division of Power to replace our power grid to the very best requirements obligatory in order that no quantity of disaster will render america defenseless by being with out power sources to energy our nation. This division is remitted to work with the EPA to enhance water and air qualities so that each neighborhood is furnished with sufficient clear contemporary water in order that one other episode of what occurred and is going on in Flint Michigan won't ever occur once more any the place in america. The roles are huge however to disregard the realities dealing with our nation at this time put america and the remainder of the world in jeopardy. These are the needs and objectives of america Division of Financial Growth.


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The ubiquitously expressed c-Abl tyrosine kinase is activated in the apoptotic response of cells to DNA damage. The mechanisms by which c-Abl signals the induction of apoptosis are not understood. Here we show that c-Abl binds constitutively to the mammalian homolog of the Schizosaccharomyces pombe Rad9 cell cycle checkpoint protein. The SH3 domain of c-Abl interacts directly with the C-terminal region of Rad9. c-Abl phosphorylates the Rad9 Bcl-2 homology 3 domain (Tyr-28) in vitro and in cells exposed to DNA-damaging agents. The results also demonstrate that c-Abl-mediated phosphorylation of Rad9 induces binding of Rad9 to the antiapototic Bcl-xL protein. The regulation of Rad9 by c-Abl in the DNA damage response is further supported by the demonstration that the interaction between c-Abl and Rad9 contributes to DNA damage-induced apoptosis. These findings indicate that Rad9 is regulated by a c-Abl-dependent mechanism in the apoptotic response to genotoxic stress.
The Schizosaccharomyces pombe checkpoint proteins spRad1, spHus1, and spRad9 are essential for cell cycle arrest in the presence of incomplete DNA replication or DNA damage (3, 4). Identification of the human homologs hRad1, hHus1, and hRad9 has provided support for conservation of this checkpoint signaling pathway (14, 29, 32, 38, 48). Also, as shown for yeast, hRad9 forms a complex with hRad1 and hHus1 (45, 50). Nuclear hRad9 is constitutively detectable as multiple phosphorylated forms (45). Other studies have demonstrated that hRad9 is phosphorylated in response to DNA damage (50) and that hRad9 localizes to extraction-resistant nuclear complexes in the presence of DNA lesions (7). Recombinant hRad9 has been shown to exhibit 3′-5′ exonuclease activity (6). Moreover, the finding that Rad9 interacts with the antiapoptotic Bcl-2 and Bcl-xL proteins has supported a role for Rad9 in the apoptotic response to DNA damage (26, 27).
The ubiquitously expressed c-Abl tyrosine kinase is activated in the response to DNA damage (23). Nuclear c-Abl interacts with the DNA-dependent protein kinase (DNA-PK)/Ku complex (17, 19). Phosphorylation of c-Abl by the catalytic subunit DNA-PKcs induces c-Abl activity (19). Other work has demonstrated that c-Abl is activated by the product of the gene mutated in ataxia telangiectasia (ATM) (5, 44). Activation of nuclear c-Abl by DNA damage contributes to induction of apoptosis by mechanisms in part dependent on the p53 tumor suppressor and its homolog p73 (2, 15, 54, 57, 58). Nuclear c-Abl also contributes to DNA damage-induced activation of the c-Jun N-terminal kinase/stress-activated protein kinase (SAPK) and p38 mitogen-activated protein kinase pathways (20, 22, 23, 37). In addition, the finding that c-Abl interacts with the Rad51 protein in response to DNA damage has supported a role for c-Abl in coordinating recombinational DNA repair with the induction of apoptosis (56).
The present studies demonstrate that c-Abl interacts with Rad9 in cells exposed to DNA-damaging agents. c-Abl phosphorylates Rad9 on Y28 in the Bcl-2 homology 3 (BH3) domain and induces binding of Rad9 to Bcl-xL. The results also demonstrate that c-Abl regulates the apoptotic response to genotoxic stress by a Rad9-dependent mechanism.
Cell culture.Human U-937 myeloid leukemia cells (American Type Culture Collection [ATCC]) and U-937 cells overexpressing Bcl-xL (U-937/Bcl-xL) (9) were cultured in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS), 100 U of penicillin/ml, 100 μg of streptomycin/ml, and 2 mM l-glutamine. 293T embryonal kidney cells (ATCC), HeLa cells (ATCC), wild-type (c-abl +/+) murine embryo fibroblasts (MEFs), c-abl −/− MEFs (23, 47), and c-abl −/− MEFs expressing c-Abl (c-abl +) (23) were grown in Dulbecco's modified Eagle's medium containing 10% FBS and antibiotics. Cells were treated with 10 μM ara-C (Sigma-Aldrich). Irradiation was performed at room temperature with a Gammacell 1000 (Atomic Energy of Canada) and a 137Cs source emitting at a fixed rate of 0.21 Gy/min. In certain experiments, cells were pretreated with 1 μM STI-571 (Novartis Pharma AG) (11) for 24 h and then exposed to ara-C or radiation.
Plasmid construction.The vector expressing green fluorescent protein (GFP)-Rad9 has been described (27). The Rad9(Y28-F) variant was generated by site-directed mutagenesis, and the mutation was confirmed by DNA sequencing. Rad9 (1-265) and Rad9 (266-391) were generated by PCR. The Rad9 cDNAs were subcloned into the pHM6 (hemagglutinin [HA] tagged; Roche Molecular Biochemicals), pLXSN (Clontech), pGEX4T-1 (Pharmacia Biotech), and pET-28a (Novagen) vectors.
Cell transfections and infections.293T cells were transiently transfected by the calcium phosphate method. The total DNA concentration was kept constant by including an empty vector. For retrovirus infections, PG13 cells (34) were transiently transfected with pLXSN-Rad9 or pLXSN-Rad9(Y28-F) by the Superfect transfection kit (Qiagen). At 48 h posttransfection, filtered supernatants with Polybrene (Sigma-Aldrich) were added to the MEFs.
Immunoprecipitation and immunoblot analysis.Cell lysates were prepared as described previously (52, 53) and cleared by centrifugation at 12,000 × g for 15 min. Soluble proteins (500 μg) were incubated with anti-Rad9 (sc-8324; Santa-Cruz Biotechnology [SCBT]), anti-c-Abl (sc-131; SCBT), anti-HA (Roche Molecular Biochemicals), or anti-GFP (Roche Molecular Biochemicals) for 2 to 6 h at 4°C, followed by incubation with protein A/G-Sepharose beads (SCBT) for 1 h. The immune complexes and cell lysates (50 μg) were subjected to immunoblot analysis with anti-Rad9 (Transduction Laboratories), anti-c-Abl (Ab-3; Oncogene Research Products), anti-HA, anti-GFP, anti-glutathione S-transferase (GST; Upstate Biotechnology Inc.), anti-P-Tyr (4G10; Upstate Biotechnology), or anti-Bcl-xL (21). The antigen-antibody complexes were visualized by chemiluminescence (NEN Life Science Products). Intensity of the signals was quantitated by densitometric scanning and analysis with the ImageQuant program (Molecular Dynamics, Sunnyvale, Calif.).
Glycerol gradient sedimentation analysis.Nuclear extracts from U-937 and U-937/Bcl-xL cells were prepared as described previously (10). Two hundred microliters of nuclear extracts was layered on a 10 to 35% glycerol gradient (5 ml) formed in a solution containing 50 mM Tris-HCl [pH 7.3], 0.1 M KCl, 0.2 mM EDTA, 10 mM β-mercaptoethanol, and 0.1% NP-40. After centrifugation at 250,000 × g for 12 h at 4°C, fractions (150 μl) were collected from the bottom of the tube. Equal volumes (20 μl) of each sample were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting with anti-c-Abl, anti-Rad9, or anti-Bcl-xL. As controls, protein standards (Sigma) were separated under identical conditions.
In vitro binding assays.Cell lysates were incubated with purified GST, GST-c-Abl SH2, and GST-c-Abl SH3 (58) in lysis buffer for 2 h at 4°C. The adsorbates were analyzed by immunoblotting with anti-Rad9 or anti-GST. For direct binding assays, GST, GST-c-Abl, and GST-c-Abl SH3 (58) were incubated with purified His-Rad9, His-Rad9 (1-265), or His-Rad9 (266-391) for 1 h at 4°C. The complexes were subjected to immunoblot analysis with anti-Rad9 or anti-His (SC803; SCBT).
In vitro kinase assays.A recombinant c-Abl protein was purified from Sf9 cells infected with a baculovirus vector expressing GST-c-Abl or GST-c-Abl(K-R). The procedure for purifying GST-c-Abl with glutathione beads was as described previously (33) and as modified by our laboratory (22). Purity of the recombinant protein was ≥90% as determined by SDS-PAGE and Coomassie brilliant blue staining. GST-c-Abl, GST-c-Abl(K-R), and c-Abl with the SH3 domain deleted (c-AblΔSH3; Oncogene) were incubated in kinase buffer (50 mM HEPES [pH 7.4], 10 mM MgCl2, 10 mM MnCl2, 2 mM dithiothreitol, 0.1 mM sodium vanadate) with purified GST-Rad9, GST-Rad9(Y28-F), GST-Rad9 (1-265), or GST-Crk (120-225) and [γ-32P]ATP (3,000 Ci/mmol; NEN Life Science Products) for 15 min at 30°C. Phosphorylated proteins were separated by SDS-PAGE and analyzed by autoradiography.
In vitro Rad9/Bcl-xL complex assays.GST, GST-Rad9, GST-Rad9(Y28-F), and GST-Rad9 (1-265) were incubated in kinase buffer with or without purified c-Abl and ATP for 30 min at 30°C and then coincubated with His-Bcl-xL (21) in lysis buffer for 1 h at 4°C. Adsorbates to glutathione beads and the residual supernatants were subjected to immunoblotting with anti-Bcl-xL.
Apoptosis assays.DNA content was assessed by staining ethanol-fixed cells with propidium iodide and monitoring by FACScan (Becton Dickinson). The numbers of cells with sub-G1 DNA were determined with a CellQuest program (Becton Dickinson).
c-Abl associates with Rad9.To investigate whether c-Abl associates with Rad9, lysates from 293T cells cotransfected with c-Abl and a HA epitope-tagged Rad9 (HA-Rad9) were subjected to immunoprecipitation with an anti-HA antibody. Analysis of the immunoprecipitates with anti-c-Abl showed binding of c-Abl and Rad9 (Fig. 1A, left). The reciprocal experiment, in which anti-c-Abl immunoprecipitates were analyzed by immunoblotting with anti-HA, confirmed coimmunoprecipitation of c-Abl and Rad9 (Fig. 1A, right). Similar findings were obtained with HeLa cells cotransfected with c-Abl and Rad9 (data not shown). To assess whether endogenous c-Abl associates with endogenous Rad9, anti-c-Abl immunoprecipitates from U-937 cells were subjected to immunoblotting with anti-Rad9. The results demonstrate constitutive binding of endogenous c-Abl and Rad9 (Fig. 1B). Immunoblot analysis of anti-Rad9 immunoprecipitates with anti-c-Abl provided further support for constitutive binding of the endogenous c-Abl and Rad9 proteins (Fig. 1B). To determine whether DNA damage affects the binding of c-Abl and Rad9, U-937 cells were treated with ara-C or ionizing radiation (IR). Analysis of anti-c-Abl immunoprecipitates with anti-Rad9 demonstrated little if any change in c-Abl-Rad9 complexes for ara-C-treated, compared to control, cells (Fig. 1C). Similar results were obtained with IR-treated cells (Fig. 1C). To confirm the constitutive binding of c-Abl and Rad9, nuclear lysates from U-937 cells were subjected to sedimentation in a glycerol gradient. Analysis of the gradient fractions by immunoblotting with anti-c-Abl and anti-Rad9 demonstrated cosedimentation of c-Abl and Rad9 in fractions 19 to 23 (Fig. 1D). These findings collectively demonstrate that c-Abl associates with Rad9 in cells and that constitutive binding is not affected by DNA damage.
To further define the association of c-Abl and Rad9, lysates from U-937 cells were incubated with GST fusion proteins containing the c-Abl SH2 or SH3 domain. Analysis of the adsorbates with anti-Rad9 showed binding of Rad9 to GST-c-Abl SH3 but not to GST or GST-c-Abl SH2 (Fig. 2A). The identification of several potential sequences for c-Abl SH3 binding (12, 40) in the Rad9 C-terminal proline-rich region (PQPP, amino acids 331 to 334; PKSP, amino acids 334 to 337; PGTP, amino acids 353 to 356; PQGP, amino acids 376 to 379) suggested that there is a direct interaction between these two proteins. To assess involvement of the Rad9 C-terminal region in binding to c-Abl, a His-tagged Rad9 (1-265) fusion protein with a deletion of the C-terminal 126 amino acids was generated. The association of His-Rad9(full length [FL]), but not His-Rad9 (1-265), with purified GST-c-Abl indicated that c-Abl binds directly to the C-terminal region of Rad9 (Fig. 2B). In addition, the finding that GST-c-Abl SH3, and not GST, binds to His-Rad9 (266-391) provided further support for the direct binding of the c-Abl SH3 domain to the Rad9 C-terminal region (Fig. 2C).
Rad9 Y-28 is required for c-Abl phosphorylation.To assess whether c-Abl phosphorylates Rad9, we incubated purified kinase-active c-Abl, kinase-inactive c-Abl(K-R), and c-AblΔSH3 with GST-Rad9 and [γ-32P]ATP. Analysis of the products by SDS-PAGE and autoradiography showed that Rad9 is a substrate for c-Abl but not c-Abl(K-R) in vitro (Fig. 3A). The finding that phosphorylation of Rad9 by c-AblΔSH3 is decreased compared to that obtained with full-length c-Abl indicates that the binding of the c-Abl SH3 domain facilitates Rad9 phosphorylation (Fig. 3A). Rad9 contains a BH3 domain but not other BH domains that are characteristic of the Bcl-2 family (27). Conserved BH3 domains within proapoptotic proteins are critical for binding to and neutralizing the activity of anti-apoptotic Bcl-2 family members (1, 18). A Y28ELP site in the Rad9 BH3 domain is a consensus sequence for c-Abl phosphorylation (46, 60). The finding that mutation of Rad9 Y28 to F completely inhibits c-Abl-mediated phosphorylation indicates that Y28 is the major c-Abl site (Fig. 3B, left). Moreover, the finding that c-Abl phosphorylates Rad9 but not Rad9 (1-265) indicates that binding to the C terminus is necessary for phosphorylation of the Y28 site (Fig. 3B, right). As confirmation that the phosphorylation of Rad9 is mediated by c-Abl, other studies were performed in the presence of c-Abl inhibitor STI-571 (11). The results demonstrate that STI-571 inhibits phosphorylation of Rad9 (Fig. 3C). To determine whether c-Abl phosphorylates Rad9 in vivo, kinase-active c-Abl or kinase-inactive c-Abl(K-R) was coexpressed with GFP-Rad9. Analysis of anti-GFP immunoprecipitates by immunoblotting with anti-P-Tyr demonstrated c-Abl-dependent tyrosine phosphorylation of Rad9 (Fig. 3D). As a control, there was little if any c-Abl-mediated phosphorylation of the Rad9(Y28-F) mutant (Fig. 3D). These findings demonstrate that the Rad9 Y28 site is required for c-Abl phosphorylation in vitro and in vivo.
DNA damage induces tyrosine phosphorylation of Rad9 by a c-Abl-dependent mechanism.To determine whether endogenous Rad9 is phosphorylated on tyrosine in cells, anti-Rad9 immunoprecipitates were analyzed by immunoblotting with anti-P-Tyr. The results demonstrate that treatment of cells with ara-C was associated with induction of Rad9 phosphorylation on tyrosine (Fig. 4A). ara-C incorporates into elongating DNA strands, inhibits DNA replication, and induces DNA double-strand breaks (13, 31, 36). The finding that exposure of cells to IR also induces tyrosine phosphorylation of Rad9 confirmed that this response is activated by genotoxic stress (Fig. 4A). To assess whether DNA damage induces tyrosine phosphorylation of Rad9 by a c-Abl-dependent mechanism, c-abl +/+, c-abl −/−, and c-abl + MEFs (Fig. 4B) (23) were treated with ara-C. Immunoblot analysis of anti-Rad9 immunoprecipitates with anti-P-Tyr demonstrated ara-C-induced tyrosine phosphorylation of Rad9 in c-abl +/+ and c-abl + MEFs (Fig. 4C). By contrast, there was no detectable tyrosine phosphorylation of Rad9 in ara-C-treated c-abl −/− MEFs (Fig. 4C). To confirm that Rad9 is phosphorylated by c-Abl, U-937 cells were treated with STI-571 and then ara-C. Immunoblot analysis of anti-Rad9 immunoprecipitates with anti-P-Tyr demonstrated that STI-571 inhibits tyrosine phosphorylation of Rad9 (Fig. 4D, left). Similar findings were obtained with c-abl + cells (Fig. 4D, right). To determine whether Rad9 is phosphorylated on Y28 in response to DNA damage, cells expressing GFP-Rad9 or GFP-Rad9(Y28-F) were treated with ara-C (Fig. 4E). Anti-GFP immunoprecipitates were analyzed by immunoblotting with anti-P-Tyr. The demonstration that ara-C induces tyrosine phosphorylation of GFP-Rad9 but not GFP-Rad9(Y28-F) supported the specificity of the Y28 site (Fig. 4E). These findings demonstrate that Rad9 is phosphorylated on Y28 by a c-Abl-dependent mechanism in response to genotoxic stress.
c-Abl-mediated phosphorylation of Rad9 induces binding of Rad9 to Bcl-xL.Recent studies have demonstrated that Rad9 binds to Bcl-2 and Bcl-xL and that these interactions are dependent on the Rad9 BH3 domain (26, 27). To determine whether c-Abl-mediated phosphorylation of the Rad9 BH3 domain regulates the association of Rad9 and Bcl-xL in vitro, GST-Rad9 was first incubated with c-Abl and ATP. Incubation of the reaction products with His-Bcl-xL and analysis of products of adsorption to glutathione beads by immunoblotting with anti-Bcl-xL demonstrated that c-Abl induces binding of Rad9 to Bcl-xL (Fig. 5A). By contrast, c-Abl had no detectable effect on the association of Rad9(Y28-F) and Bcl-xL (Fig. 5A). To extend this analysis, GST-Rad9 was incubated with c-Abl or c-Abl(K-R) and then assayed for binding to Bcl-xL. The results show that c-Abl, and not c-Abl(K-R), induces the formation of Rad9-Bcl-xL complexes (Fig. 5B). As an additional control, GST-Rad9 (1-265) was incubated with c-Abl or c-Abl(K-R). In agreement with the finding that Rad9 (1-265) is not phosphorylated by c-Abl (Fig. 3B), the binding of Rad9 (1-265) to Bcl-xL was unaffected by c-Abl or c-Abl(K-R) (Fig. 5B). In in vivo studies, the association of Rad9 and Bcl-xL was induced in cells expressing c-Abl (Fig. 5C). In agreement with the in vitro findings, c-Abl(K-R) had no detectable effect on the association of Rad9 and Bcl-xL in cells (Fig. 5C). Moreover, c-Abl had no detectable effect on the association of Rad9(Y28-F) and Bcl-xL in cells (Fig. 5D).
Whereas the results demonstrate that Rad9 is phosphorylated by c-Abl in response to DNA damage and that c-Abl induces the binding of Rad9 and Bcl-xL, studies were performed on U-937 cells expressing Bcl-xL. Analysis of nuclear lysates from U-937/Bcl-xL cells demonstrated cosedimentation of Rad9 and Bcl-xL (Fig. 6A). Constitutive binding of Rad9 and Bcl-xL was confirmed by immunoblot analysis of anti-Rad9 immunoprecipitates with anti-Bcl-xL (Fig. 6B). Importantly, the constitutive association of Rad9 and Bcl-xL was increased by ara-C treatment (Fig. 6B). Similar results were obtained with IR-treated cells (Fig. 6B). To further define the role of c-Abl in regulating the interaction of Rad9 and Bcl-xL, lysates from ara-C-treated c-abl +/+, c-abl −/−, and c-abl + cells were subjected to immunoprecipitation with anti-Rad9. Analysis of the precipitates by immunoblotting with anti-Bcl-xL demonstrated that DNA damage-induced formation of Rad9-Bcl-xL complexes is mediated by a c-Abl-dependent mechanism (Fig. 6C). In agreement with these results, pretreatment of c-abl + cells with STI-571 blocked ara-C-induced formation of Rad9-Bcl-xL complexes (Fig. 6D). These findings collectively indicate that c-Abl-mediated phosphorylation of the Rad9 BH3 domain induces binding of Rad9 to Bcl-xL.
c-Abl potentiates Rad9-mediated apoptosis.To further assess the functional significance of the interaction between c-Abl and Rad9, 293T cells were transfected to express Rad9 and c-Abl or c-Abl(K-R) (Fig. 7A). The results demonstrate that c-Abl, and not c-Abl(K-R), potentiates the effects of Rad9 on the induction of 293 cell apoptosis (Fig. 7B). To extend this analysis, c-abl +/+, c-abl −/−, and c-abl + MEFs were infected with an empty retrovirus vector or one that expresses Rad9 (Fig. 7C). The apoptotic response of c-abl +/+ and c-abl + MEFs to Rad9 was more pronounced than that obtained with c-abl −/− cells (Fig. 7C; 0 h). c-abl +/+ and c-abl + MEFs expressing Rad9 also responded to ara-C treatment (Fig. 7C, 12 and 24 h) with a greater induction of apoptosis than that found with c-abl −/− cells expressing Rad9 (Fig. 7C). Moreover, expression of Rad9(Y28-F) was associated with attenuation of the apoptotic responses found with Rad9 (Fig. 7C). In agreement with these results, pretreatment of c-abl + cells with STI-571 inhibited Rad9-induced apoptosis (Fig. 7D). These results support a functional interaction between c-Abl and Rad9 that contributes to the induction of apoptosis.
Role of c-Abl in the apoptotic response to genotoxic stress.The mechanisms by which DNA damage is converted into intracellular signals that control cell behavior are largely unknown. Certain insights have been derived from the finding that the c-Abl tyrosine kinase is activated by agents that arrest DNA replication (e.g., ara-C) or induce DNA lesions (e.g., IR) (20, 23). The available evidence indicates that c-Abl functions in diverse aspects of the DNA damage response, including the activation of cell death signals. For example, transient transfection studies have demonstrated that wild-type c-Abl, but not kinase-inactive c-Abl(K-R), induces an apoptotic response (55). In addition, cells that stably express c-Abl(K-R) exhibit resistance to induction of apoptosis by ara-C and IR (16, 55). These studies and the present work, which employs ectopic expression to assess function, can result in overinterpretation of findings. Similar results have nonetheless been obtained with c-abl −/− fibroblasts, although the apoptosis-resistant phenotype is more pronounced in cells expressing c-Abl(K-R) than in c-Abl null cells (16, 55). These findings indicate that a redundant function, perhaps mediated by the related Arg kinase (30), is inhibited by the dominant-negative effect of c-Abl(K-R).
Exposure of diverse mammalian cells to ara-C, IR, and other DNA-damaging agents is associated with activation of c-Abl and SAPK (8, 20, 23, 49). The demonstration that c-Abl-deficient cells exhibit a defective SAPK response to DNA damage has supported a role for c-Abl in regulation of the SAPK pathway (20, 23). More recent work has shown that c-Abl interacts directly with MEK kinase 1, an upstream effector of the SEK1→SAPK pathway, in response to DNA damage (22). In agreement with these findings, other studies have demonstrated that transient overexpression of activated forms of Abl stimulates SAPK activity (39, 41, 42). The functional significance of c-Abl-induced SAPK activation in the DNA damage response is supported by the demonstration that SAPK translocates to mitochondria and associates with the Bcl-xL protein (24). SAPK phosphorylates Bcl-xL on Thr-47 and Thr-115 and thereby contributes to the induction of apoptosis (24). In the present studies, we provide evidence that c-Abl also targets Bcl-xL by a mechanism involving activation of Rad9.
c-Abl phosphorylates Rad9.In human cells, hRad9 forms a ternary complex with hRad1 and hHus1 (45, 50). The findings that hRad9 and hHus1 interact with the proliferating nuclear cell antigen suggested that the Rad9 complex contributes to the coordination of cell cycle progression, DNA replication, and DNA repair (28). Although few insights regarding the regulation of hRad9 are available, multiple phosphorylated forms of hRad9 have been detected in complexes with hRad1 and hHus1 (45). The kinases responsible for the modification of hRad9 and the functional significance of this regulation are unknown.
The present studies show that c-Abl binds directly to Rad9 and phosphorylates Rad9 on Y28 in vitro and in cells. The results also demonstrate that Rad9 is subject to c-Abl-mediated phosphorylation on Y28 in cells treated with genotoxic agents. Moreover, treatment of cells with 1 μM STI-571 partially inhibited phosphorylation of Rad9 on tyrosine. In agreement with these findings, activation of c-Abl in the DNA damage response is partially inhibited with 1 μM STI-571 and completely inhibited with 10 μM STI-571 (data not shown). Thus, inhibition of DNA replication by ara-C treatment or induction of DNA lesions by IR exposure induces tyrosine phosphorylation of Rad9 by a c-Abl-dependent mechanism. Rad9 contains a motif of 15 amino acids near the N terminus (amino acids 15 to 30) that includes conserved residues found in BH3 domains of proapoptotic Bcl-2 family members (27). The BH3 domain-only proteins function as transdominant inhibitors by binding to antiapoptotic members of the Bcl-2 family and blocking their survival function (1, 18). Notably, Y28 resides in the Rad9 BH3 domain. Moreover, the Rad9 Y28 site is unique to BH3 consensus sequences found in other proapoptotic members of the Bcl-2 family. These findings support a model in which c-Abl-mediated phosphorylation of the Rad9 BH3 domain is selective for this proapoptotic member of the Bcl-2 family.
c-Abl regulates the interaction between hRad9 and Bcl-xL . We have previously shown that the Rad9 BH3 domain regulates the interaction of Rad9 with the antiapoptotic Bcl-2 and Bcl-xL proteins (27). The present results demonstrate that c-Abl-mediated phosphorylation of Rad9 on Y28 induces the interaction of Rad9 and Bcl-xL. In vitro studies show that Rad9 associates constitutively with Bcl-xL and that c-Abl stimulates this interaction. By contrast, c-Abl had no effect on constitutive binding of Rad9(Y28-F) to Bcl-xL. In transfected cells, c-Abl also increased the interaction of Rad9, but not of Rad9(Y28-F), with Bcl-xL. In agreement with these results, exposure of cells to agents, such as ara-C and IR, that activate c-Abl (20, 23) was associated with binding of Rad9 to Bcl-xL. By contrast, results obtained with c-Abl-deficient cells or after treatment of wild-type cells with STI-571 indicate that c-Abl has little if any effect on the binding of Rad9 to Rad1 and Hus1 in response to DNA damage (data not shown). The finding that DNA damage induces the interaction between Rad9 and Bcl-xL in c-abl +/+ and c-abl +, but not in c-abl −/−, cells further indicated that this response is c-Abl dependent.
Nuclear magnetic resonance and X-ray crystallographic analysis of the Bcl-xL monomer has demonstrated that the α-helices of BH1 to -3 form a hydrophobic pocket (35). The BH3 amphipathic α2-helix of proapoptotic family members is essential for binding to the pocket and for inhibition of Bcl-xL function (43, 51, 59). The present results support a model in which modification of the Rad9 BH3 α-helix by c-Abl-mediated phosphorylation is associated with a structural change that facilitates binding to the Bcl-xL hydrophobic pocket. Other studies have demonstrated that c-Abl functions upstream of SAPK-mediated phosphorylation of Bcl-xL (24). SAPK phosphorylates Bcl-xL on (i) Thr-47, which resides in the loop between the α1- and α2-helices, and (ii) Thr-115, which is adjacent to the α3-helix (24). The demonstration that the Bcl-xL(A-47,-115) mutant is more effective than wild-type Bcl-xL in blocking apoptosis indicates that phosphorylation of Thr-47 and Thr-115 is involved in regulating Bcl-xL function (24). These findings suggest that modifications of both the Rad9 BH3 domain by c-Abl and of Bcl-xL directly by SAPK are involved in regulating the function of Bcl-xL in protecting against apoptosis.
c-Abl potentiates the proapoptotic function of Rad9.The results of the present study demonstrate that Rad9-induced apoptosis is potentiated by expression of c-Abl and not by the kinase-inactive c-Abl(K-R) mutant. As c-Abl interacts with multiple targets that may contribute to DNA damage-induced apoptosis (25), we performed studies to assess the contribution of the c-Abl-Rad9 interaction to this response. The demonstration that expression of Rad9 and that of Rad9(Y28-F) in cells are associated with similar levels of apoptosis indicated that mutation of Y28 has no apparent effect on Rad9's basal proapoptotic function. By contrast, the results show that DNA damage-induced apoptosis is potentiated by expression of Rad9 and not by that of Rad9(Y28-F). In addition, Rad9-mediated induction of apoptosis in response to DNA damage was attenuated in c-Abl-deficient cells. These findings collectively support a model in which c-Abl-mediated phosphorylation of Rad9 in the DNA damage response contributes to the binding of Rad9 to Bcl-xL and thereby to potentiation of Rad9-induced apoptosis.
ACKNOWLEDGMENTSThis work was supported by grants CA29431 and CA55241 awarded by the National Cancer Institute.
We thank R. Ren and D. Baltimore for providing the c-abl −/− and c-abl + fibroblast cell lines. We appreciate the helpful discussions with Jun-ichi Sawada and the technical assistance of Kamal Chauhan.
- Received 19 November 2001.
- Returned for modification 8 January 2002.
- Accepted 12 February 2002.
- Copyright © 2002 American Society for Microbiology

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Early results of a trial comparing high-dose with conventional chemotherapy for women with breast cancer show that there are no apparent differences in survival or in the rate of relapse, according to research conducted in the UK, Ireland, New Zealand, and Belgium. Speaking at the European Society for Medical Oncology Congress in Nice, France today (21 October 2002), Dr John Crown from St Vincent's University Hospital, Dublin, Ireland, said, however, that new drugs that target the molecules associated with cancer, given in conjunction with chemotherapy, offer promising prospects for clinical trials.
Breast cancer is the most common cause of death from cancer in women in most Western countries and leads to untold distress for their families. Whilst not all patients die from their disease, patients whose cancer also involves the lymph nodes have a poor prognosis. Hormone therapy and chemotherapy have helped a number of patients and reduced the risk of relapse over the last two decades.
One treatment strategy for women with breast cancer has been an intense programme of chemotherapy, in combination with the patient's own bone marrow cells. High-dose chemotherapy, however, produces severe side-effects, such as prolonged suppression of blood cells, infection and bleeding. Researchers have focused their studies on high-dose chemotherapy both in patients whose cancer has spread to other organs, as well as high-risk patients at an early stage of the disease. "There were great expectations for high-dose chemotherapy, but evidence from a number of trials show that there are no real bene
Contact: Gracemarie Bricalli
European Society for Medical Oncology


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In these unprecedented times, we have come together united as a faith and learning community. In the weeks to come, we will share positive stories from students, parents, and staff – stories of who we are as a Catholic community - on our website and through our social media platforms.
We invite you to share your own stories of faith and learning at home with us. Please use the hashtags #ABLearnAtHome, #GSACRDProud, and #GSACRDgrace when telling your stories.
In our third community story, students and staff share what hope means to them. They also describe some personal challenges they have faced and how those challenges have helped them become more hopeful, especially in light of COVID-19 and this Easter and Spring season.
Watch the video below.
Previous community stories about faith and learning at home
- Week 2 (April 9, 2020): What we like most about online learning and Easter
- Week 1 (April 3, 2020): The importance of faith and prayer


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Lincoln University is a higher education institution located in Alameda County, CA. In 2016, the most popular Bachelor's Degree concentrations at Lincoln University were Diagnostic Medical Sonography & Ultrasound Technician (13 degrees awarded) and General Business Administration & Management (8 degrees).
In 2017, 123 degrees were awarded across all undergraduate and graduate programs at Lincoln University. 46.3% of these degrees were awarded to women, and 53.7% awarded men. The majority of degree recipients were asian (12 degrees), 3 times more than then the next closest race/ethnicity group, black or african american (4 degrees).
The median undergraduate tuition at Lincoln University is $10,440, which is $-3,560 less than the national average for Special Focus Institutions ($14,000).


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Samantha Fazio, PT, DPT, PHC
PT School and Year Graduated:
Temple University, Class of 2017
Certifications and professional accomplishments:
Pelvic Health Certification – through Evidence in Motion
E&A Therapy Clinical Achievement Award - for dedication in the pursuit of professional excellence
Temple University Hyman Dervitz Scholarship Award - in recognition of academic excellence and demonstrating collaborative leadership and social conscience.
What do you like most about being a PT?
As a pelvic health physical therapist, it is so rewarding to help people get their quality of life back in regard to basic human needs. I also love that I am afforded time to get to know my patients on a more personal level than most other healthcare providers. PT isn't always easy so I try to make each session fun and full of laughter.
Something personal:
In my spare time, I enjoy spending time with family and friends especially over good food. I also love to see shows on Broadway, read, cook, and do anything on or near a body of water!← Back to Staff


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The downside to their partner dashboard is there is no associate tracking I.d.s for associates. For instance, if an associate composed a critique and created a YouTube online video assessment advertising your product, they won't know which traffic resource (blog site or even YouTube) created the sale. This is actually why tracking IDs are vital for partners. Having said that, for the Costs pricing, I was actually anticipating the capacity for associates to include monitoring I.d.s to their links. Straightforward platform to sell electronic or even physical items onlineSales funnel functionalities: purchase bumps & one click upsells to improve order valueAdvanced statistics stating like Lifetime ValueCoupon regulations are effortless to create (% or even $ off)Repayment Versions: single, registrations (payment regularity control), remittance plans, trialsLanding web page A/B crack testingKnowledge foundation has SamCart instructions resourcesCheckout page pushcart desertion capabilitiesBuilt-in Dunning Device (Range Program)Pull and also Fall Publisher for have a look at pagesOnly sustains Red stripe and PayPalMonthly pricing beginning at $49 every monthNo electronic sales tax obligation computation built-inAffiliate system doesn't have tracking IDs partners can useSamCart provides 3 regular monthly rates programs.$49/month for the Introduce Strategy$99/month for the Grow Planning$199/month for the Scale PlanThe is fit for internet business people who are actually launching their first electronic item or even bodily item.
These components are actually unlocked in the Grow as well as Scale strategies. The includes what the Launch Planning does not have. Features such as client administration, membership fee tips, accelerated subscription terminations, and even more. The includes Advanced SamCart's functions. It consists of all the features on call in the Launch as well as Develop plan in addition to their built-in dunning tool, their Partner Facility, cart abandonment, integrated AB screening, CRM integrations, webhooks as well as priority email & chat assistance.
Each pricing plan possesses a free test option as well as a money-back assurance. Since our company've dealt with SamCart's standard and also accelerated features permit's find out if it's is the appropriate buying pushcart answer for.I have actually boiled this SamCart customer review to two cases when I will advise you utilize SamCart and also one when I definitely would not – straightforward.
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You have a subscription-based businessThe Range Plan is actually worth it if you have a membership items due to their dunning system. You would like to supply your affiliates along with various sorts of tracking links to advertise your product(s)SamCart has a wonderful associate function for item proprietors that possess existing material that directs leads to their products.
Any kind of page that has your material advertising set up. You'll be actually capable to establish these webpages as partner links your associates can easily utilize (besides the primary product web page affiliate hyperlink). You would like to one more month-to-month registration chargeSamCart's rates choices are good, however when reviewed to ThriveCart, which carries out a lot of what SamCart's Grow and also Scale planning may do, I will highly recommend paying for $495 single (assessment hyperlink) for ThriveCart if you desire to avoid the month to month charge.
Although SamCart is actually a significant month-to-month investment, no person can easily refuse that it is actually a highly effective system for online marketing experts (specifically their Incrustation Strategy). Apart from its state-of-the-art components, their simple interface (and brand new drag & reduce publisher) are going to let you create stunning check out web pages that will certainly separate you from your competitors.
ThriveCart is actually giving them an actual jog for their money for check out page software (they have a drag-and-drop cart webpage home builder as well). Having actually utilized each platforms, I would certainly encourage the SamCart's Scale Program at $199 each month and also ThriveCart as opposed to SamCart's Introduce or Grow Plan (unhappy Brian M.). offer SamCart a shot, the free of cost test lasts for 2 week – enough to observe if their shopping cart suits effectively with your online business!If you have concerns, leave them in the remarks listed below.
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They supply a free profile strategy, however there is actually certainly not a great deal of modification or even sophisticated channel attributes like upsells and also purchase bumps that come along using it. is SamCart's # 1 shopping pushcart competitor because they provide one-time costs for much of the exact same functions. Because TC is actually a life-time price, it doesn't use a free of charge trial.is a fantastic appearing buying pushcart if you desire to have a popup have a look at process that combines effortlessly with your website – Cheap Landing Page Software For Sale Near Me.is a sound platform I have actually however, to assess fully, yet I have listened to benefits coming from users.


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iPath disrupted all markets with their RPA (Robotic Process Automation) system. Simply put, they made it easier for anyone to automate the tedious, repetitive tasks we all struggle with in the workplace.
And their history is inspiring. They started back in 2005, and haven't stopped growing since.
"We started in 2005 as a 10-people team based in Bucharest, led by Daniel Dines. In the beginning, we outsourced automation libraries and software to some of the world's biggest companies. Fast-forward to 2015. We launched free courses for UiPath Academy and started teaching RPA to over 35,000 now certified developers, propelling our rocketship growth.
Today, we have 5,000+ enterprise customers and have raised over $920M to a valuation of $7 billion. And the best is yet to come."
As you know, UiPath is still going strong.
And they have big plans for the future.
"Bill Gates used to talk at Microsoft about a computer in every home. I want a robot for every person." - Daniel Dines, UiPath CEO
Bloomberg stated that they have insider information about UiPath's plans to raise even more funds, totalling their valuation at $10 billion, but the company has refused to comment on the matter.
Whether or not that's what they're planning to do, you should know it isn't far from what they achieved last year. In April of 2019, UiPath closed a financing round totaling $568 million from Coatue Management, Dragoneer Investment Group, Wellington Management and others.
The current economic landscape benefits companies like UiPath, so raising capital for rapid growth makes sense. In the wake of the CoVid crisis, many companies sought automation solutions to cut costs and stay afloat with diminishing demand for their products.
But it's not like UiPath was doing bad in any way. Some of their clients include the US Navy, The CIA, and McDonald's.
According to their own website: "UiPath is the fastest-growing enterprise software company in history. The employee base grew to over 2,900 employees today, across 25+ offices. In 2019 UiPath was named a leader in the Gartner Magic Quadrant for Robotic Process Automation Software." On top, they have a 4x growth year by year.
So you can see why investors might as well be interested in another successful round.
Especially considering how their market is expected to grow.
But to dig deeper into that…
What is Robotic Process Automation?
RPA is a subset of Business Process Automation that creates metaphorical bots with specific tasks. More often than not, you'll be able to use them to emulate a human doing repetitive tasks.
Broadly, this is what RPA bots can do:
- Log into apps
- Connect with APIs
- Do data processing
- Move files around
- Extract content from documents
- Scrape the web for data
- Read and write databases
And a lot of other stuff.
UiPath rose to prominence in this medium due to their versatile framework for developing bots. On top, as you can guess, there's no shortage of potential clients. Any company can use the RPAs to increase efficiency and productivity.
But UiPath is not the only player in town.
Blue Prism Group is a big name in the industry too, and they went public in 2016. Softomotive Ltd is a smaller, UK-based company that was acquired by Microsoft recently.
So there is some wiggle room in the industry.
But so far, UiPath has dominated the scene.
Now we bring it back to you.
Could your work benefit from something like UiPath?
Let us know down below!


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Turkish occupation works in the construction of the wall in the late night hours, and from time to time and not on a daily basis, sources pointed out that the occupation resorted to it after the reactions of this wall and anxious not to draw attention to it.
(ANHA) correspondent, citing sources, said the wall had reached 3,600 meters. Sources said that the occupation will build a 75 km wall between Afrin and Syrian territory.
The latest Turkish work to build the wall continues in the villages of Basala and Kemar, which belong to Sherawa.
The partition wall has been built in Gilbera village and Turkish occupation army approached to Basala village, as adjacent, as well as continue to build in the village of Marimin in Shara district.
The construction of the wall in some villages in Sherawa district was stopped as a result of the military operations carried out by the Afrin Liberation Forces in that area.
The Turkish occupation demolished dozens of civilian homes to build this wall, and this was seen with the naked eye in the village of Gilbera. The Turkish occupation also cut hundreds of olive and forest trees in order to build its bases and this wall.


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Know with whom you share the best and worst relations with, based on your Zodiac Sign.
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Cross-Border Activities: Benefits of Filing "Protective" Returns
With the increasing focus by the Internal Revenue Service (IRS) on offshore activities, it is important to understand the tax return and information return filing obligations that may be associated with such activities. Because application of many of these obligations is not entirely clear in certain common situations, many taxpayers (including non-U.S. persons who might not otherwise consider themselves U.S. taxpayers) may be well-advised to resolve any reasonable doubt they might have in favor of filing. Although this sometimes places significant burdens on those taxpayers, the potential advantages to making these filings are likely to outweigh the burdens.
Uncertain Application of Tax and Information Return Filing Obligations
U.S. citizens, U.S. green card holders and U.S. corporations are clearly required to file a U.S. income tax return each year if their income meets the filing threshold. Less clear is whether such U.S. persons must file "information returns" for activities in which they may be involved outside of the United States, and whether non-U.S. persons who have some U.S. business-related activity are required to file U.S. income tax returns. Some examples of these situations are as follows.
- U.S. persons involved in funding a foreign arrangement (such as a will or a retirement plan) may not readily be able to determine whether the arrangement constitutes a trust for U.S. tax purposes. Should they file additional information returns (Forms 3520 and/or 3520A) to report a foreign trust?
- A U.S. person with an ownership interest in a foreign corporation. Should he/she file an information return to disclose that ownership?
- A non-U.S. person may be involved in conducting some business-related activity in the U.S., but it is not clear whether that activity rises to the level of engaging in a U.S. trade or business. Should that person file U.S. returns?
The short answer is – when not entirely clear, the taxpayer (or potential taxpayer) is probably better off making a "protective" filing of tax returns or information returns. The primary reason is that if the IRS later concludes that such a return is required, then a protective filing of the return can preserve numerous benefits for the taxpayer which would not have been available had the taxpayer not made such a protective filing.
Reserve Benefits of Otherwise Deductible Items
Non-U.S. persons engaged in a trade or business in the U.S. are required to file a U.S. tax form even if there is no income generated from that trade or business. For individuals, the required form is called Form 1040NR (U.S. Nonresident Alien Income Tax Return). For foreign corporations, the appropriate tax return form is the Form 1120F (U.S. Income Tax Return of a Foreign Corporation). Under IRS regulations, failure to file such a return within 16 months after its due date allows the IRS to deny any deductions from adjusted gross income in determining the non-U.S. taxpayer's tax liability. However, it can be difficult to make a definite determination as to whether a non-U.S. person's activity in the U.S. rises to the level of "engaged in a trade or business within the United States." When there is lack of certainty, the non-U.S. person can preserve the right to receive benefits of otherwise deductible items by timely filing a protective return. The IRS has specifically recognized that a pro forma "protective" tax return that shows no income effectively connected with the taxpayer's U.S. trade or business, when accompanied by a statement that the return is being filed to preserve the taxpayer's right to claim applicable deductions, is sufficient to satisfy the tax return filing requirement.
Get the Statute of Limitations Running
The statute of limitations for assessing taxes generally determines the length of time that the IRS is allowed to propose adjustments to a filed tax return. If the statute of limitations has expired with respect to a particular tax year, then the IRS is precluded from making such adjustments for that year. Generally, the statute of limitations for tax returns is three years starting from the date of filing or the date it was due (if later). The statute can be extended to six years if there is a "substantial understatement" of income. If a taxpayer never files a tax return with respect to a given taxable year, effectively there is no statute of limitations on the IRS's right to assess additional taxes for that year. Regardless of the length of the statute of limitations, therefore, it is imperative to get the statute of limitations running.
The statute of limitations does not begin to run until a tax return is filed. Moreover, if certain information returns reporting transfers or ownership of assets outside the United States are required parts of the tax returns, then the statute of limitations does not begin to run until the required information returns are filed, because the tax return otherwise is considered incomplete. Sometimes, it is not entirely clear whether an information return is required to be filed, especially when foreign arrangements are involved. For instance, a U.S. person who has an interest in or makes a contribution to a foreign trust is required to file Form 3520 (Annual Return To Report Transactions With Foreign Trusts and Receipt of Certain Foreign Gifts) and/or Form 3520-A (Annual Information Return of Foreign Trust With a U.S. Owner). Unfortunately, it may not be easy to determine whether some of the foreign arrangements constitute a trust for U.S. tax purposes. A foreign estate planning arrangement or a foreign foundation may be deemed a trust for these purposes.
If there is a lack of clarity regarding the status of a foreign trust, then a taxpayer probably should take a conservative approach and make a protective filing of the IRS forms. Filing the forms can at least get the statute of limitations on the returns to begin running.
Likewise, if a U.S. person has certain involvement with or ownership interest in a foreign corporation, then that U.S. person may be required to disclose that involvement or ownership on a Form 5471 (Information Return of U.S. Persons With Respect to Certain Foreign Corporations). Again, if the facts are not entirely clear as to the requirement of filing such an information return, a protective filing should be considered to get the statute of limitations running.
Avoid Failure to File Penalties
Another advantage of making a protective filing is to avoid failure to file penalties. For example, if the IRS later determines that a foreign arrangement constitutes a trust, and Form 3520 and/or Form 3520-A should be filed, then failure to file the return can subject the taxpayer to severe penalties. If a required Form 3520 is not timely filed, then the initial penalty is $10,000 or 35 percent of the value of the trust assets in question, whichever is greater. If a required Form 3520-A is not timely filed, then the initial penalty is $10,000 or 5 percent of the gross value of the portion of the trust owned by the U.S. person, whichever is greater. Similarly, if a taxpayer is required to file Form 5471 and fails to do so, the failure to file penalty can be $10,000 for each year for each Form 5471 not filed.
Any Downside?
Of course, making a protective filing of the returns can be expensive. In addition, if the return is filed inaccurately, then there might be penalties. However, in most cases, the benefit of filing a protective return easily outweighs the downside of filing an inaccurate return when the IRS later concludes that a return should have been filed.


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Tax laws can be exceedingly complicated, particularly those governing businesses and charities. In most cases, if you have a legal question or dispute involving tax laws, you'll need to hire a tax attorney for assistance. Many people feel more comfortable working with a Christian tax attorney for major projects, as this can help prevent conflicts of interest during the process.
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If you're a Lewisham parent, wondering what's happening to your appeal regarding secondary school places, I can tell you.
(Ok, I know none of you are, but just imagine...)
The Royal Mail has taken about two weeks to deliver a packet of appeals from the Town Hall in Catford to the chair of the appeals panel, about a mile away.
How do I know? I was behind him in the queue at the delivery office as he was making his complaint.
I was there to pick up a recorded post item, but if I had wanted to make a similar point, I'd have been mentioning the birthday present that was posted - in a Post Office, from the custom 'stamp' - first class on 21st April in Nottingham which wasn't delivered here until 5th of May.