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Acetylation of the ε-amino group of lysine residues, or Nε-lysine acetylation, is an important post-translational modification known to occur in histones, transcription factors and other proteins. Since 1995, dozens of proteins have been discovered to possess intrinsic lysine acetyltransferase activity. Although most of these enzymes were first identified as histone acetyltransferases and then tested for activities towards other proteins, acetyltransferases only modifying non-histone proteins have also been identified. Lysine acetyltransferases form different groups, three of which are Gcn5/PCAF, p300/CBP and MYST proteins. While members of the former two groups mainly function as transcriptional co-activators, emerging evidence suggests that MYST proteins, such as Esa1, Sas2, MOF, TIP60, MOZ and MORF, have diverse roles in various nuclear processes. Aberrant lysine acetylation has been implicated in oncogenesis. The genes for p300, CBP, MOZ and MORF are rearranged in recurrent leukemia-associated chromosomal abnormalities. Consistent with their roles in leukemogenesis, these acetyltransferases interact with Runx1 (or AML1), one of the most frequent targets of chromosomal translocations in leukemia. Therefore, the diverse superfamily of lysine acetyltransferases executes an acetylation program that is important for different cellular processes and perturbation of such a program may cause the development of cancer and other diseases.
The question of how protein functions are regulated in vivo has been and remains a central issue in studies of various biological processes. Acetylation of the ε-amino group of lysine residues, or Nε-lysine acetylation, has recently emerged as an important covalent post-translational modification for regulating protein functions (1–7). Lysine acetylation has been mainly found in eukaryotic cells, but more recently also in archaea and eubacteria (8–11). This type of modification ought to be distinguished from acetylation of the α-amino groups of N-terminal residues, or Nα-terminal acetylation, which occurs in many eukaryotic proteins (12). Nα-terminal acetylation mainly occurs in a co-translational manner and is generally irreversible (12), whereas lysine acetylation is a reversible post-translational process. The dynamic equilibrium of lysine acetylation in vivo is governed by the opposing actions of acetyltransferases and deacetylases. The very first direct links of histone acetyltransferase (HAT) and histone deacetylase to transcriptional co-regulators, made in 1996 (13,14), initiated a gold rush to identify proteins with such enzymatic activities. It is now clear that there are three major classes of histone deacetylases (15–19). In comparison, proteins with HAT activity are more diverse (4,6,20–22). Some known HATs also actetylate other proteins (20,23). Using different protein substrates, several proteins have recently been found to possess intrinsic lysine acetyltransferase activity (24–27). Interestingly, at least two of them are unable to acetylate histones. Since acetyltransferases modifying non-histone proteins have begun to emerge, the generic term 'lysine acetyltransferases (LATs)' is used hereafter to refer to enzymes that are able to acetylate specific lysine residues within histones and/or other proteins. In what follows, I will first list various types of proteins modified by lysine acetylation and present an overview of different groups of LATs, with a special focus on the MYST family of acetyltransferases to illustrate how similar catalytic domains are used for different functional purposes in various eukaryotic organisms. I will then discuss how activities of LATs are regulated, what determines their substrate specificity and how lysine acetylation affects protein function. Evidence for the involvement of aberrant HATs in human leukemia will be described at the end, to conclude that LATs are potential molecular targets for therapeutic intervention of leukemia and other diseases caused by abnormal lysine acetylation.
The occurrence of acetyllysine in histones was first discovered in the 1960s (28–30). Core histone proteins were found to be acetylated at the ε-amino nitrogen of specific lysine residues located in the N-terminal tails (reviewed in 31). Importantly, histone acetylation appeared to be associated with active chromatin (28). In 1979, lysine acetylation of high mobility group (HMG) proteins was observed (32). In 1987, Lys40 of α-tubulin from Chlamydomonas was identified as the acetylation site (33). A decade later, in 1997, the tumor suppressor protein p53 was discovered to be acetylated at specific lysine residues in its C-terminal regulatory domain (34). Together with the finding that HATs acetylate the general transcription factors TFIIE and TFIIF in vitro (35), the unexpected discovery of p53 acetylation revealed that HATs are also able to acetylate non-histone proteins, thereby unleashing numerous studies aimed at assessing lysine acetylation in various proteins. It is well recognized now that this type of modification occurs much more widely than anticipated several years ago. As listed in Table Table1,1, over 30 DNA-binding transcription factors have been found to be Nε-acetylated (20,23,36). At least five transcriptional co-regulators are known to be similarly modified (37–41). Reminiscent of autophosphorylation found with protein kinases, several HATs are autoacetylated (42–45). Autoacetylation of PCAF (p300/CBP-associated factor) is important for nuclear localization (46) and acetylation of the general transcription factor TFIIB by itself promotes TFIIF association and transcriptional activation (26). In addition, the chromatin remodeler Brm is acetylated at a lysine residue (47). Although most known Nε-acetylated proteins are histones and transcriptional regulators (Table (Table1),1), this modification also occurs in other cellular proteins, including MCM3 (minichromosome maintenance 3) (24), DNA metabolic enzymes (48–50), the signaling regulator Smad7 (51) and α-tubulin (33). Just as in cellular proteins, Nε-acetylation has been found in viral proteins, including the HIV TAR RNA-binding protein Tat, adenoviral oncoprotein E1A and polyomavirus large T antigen (52–55). Moreover, lysine acetylation is not just unique to eukaryotic and viral proteins. The archaeal architectural protein Alba is Nε-acetylated and this modification blocks oligomerization and DNA binding (8,9). Acetylation of Lys609, a catalytic residue of acetyl-CoA synthetase from the enterobacterium Salmonella enterica, inhibits the enzymatic activity in vivo (10,11). This residue is invariant among a family of AMP-forming enzymes from prokaryotes and eukaryotes, so acetylation may modulate the function of these enzymes (10,11). More amazingly, acetylation of Lys92 dramatically up-regulates the function of Escherichia coli chemotaxis regulator CheY (56,57). Therefore, lysine acetylation has emerged as a general post-translational modification that regulates functions of various cellular and viral proteins.
In 1995, yeast HAT1 (histone acetyltransferase 1) (Table (Table2)2) was identified as the first HAT (58). Although HAT1 was considered to be mainly localized in the cytoplasm to acetylate nascent histones for deposition, recent studies indicate that this acetyltransferase also exists in the nucleus to regulate gene silencing (59). The importance of HATs in gene regulation began to be widely considered in 1996 when HAT activity was shown to be intrinsic to several known transcriptional co-activators such as Gcn5 (general control non-derepressible 5) (13), PCAF (60), p300 (E1A-associated 300 kDa protein) (43), CBP (CREB-binding protein) (43,44) and TAFII250 (TBP-associated factor of 250 kDa) (61). Subsequently, additional proteins were shown to possess HAT activity (Table (Table2).2). As listed in Table Table2,2, these proteins include the nuclear receptor co-activators SRC-1 (steroid receptor coactivator 1) and ACTR (activator of retinoid receptor, also known as AIB1 for amplified in breast cancer 1) (62,63), the transcriptional co-activator CIITA (major histocompatibility class II activator) (64), the DNA-binding transcription factor ATF2 (65), the transcriptional mediator Nut1 (66), the transcription initiation factor TFIIIC (67), the transcription elongation factor Elp3 (68), the yeast protein Hpa2 (69), CDY (chromodomain Y) and its homolog CDYL (CDY-like) (70) and the MYST family of proteins (20,22,71). p300, CBP and PCAF also acetylate transcription factors and other proteins (20,23). Some HATs display autoacetylating activity (42–45). TFIIB acetylates itself, but does not possess any detectable HAT activity (26,69). MCM3AP (MCM3 acetylating protein), Eco1 (establishment of cohesion 1) and ARD1 (a known Nα-acetyltransferase) are three other LATs that were not identified first as HATs (24,25,27). Quite interestingly, acetyl-CoA synthetase is responsible for the acetylation of CheY in E.coli (56,57).
According to sequence similarity, known LATs can be organized into different groups (Table (Table2).2). As one major group of nuclear HATs, the Gcn5/PCAF family is composed of Gcn5, PCAF and related proteins. As illustrated in Figure Figure1A,1A, yeast Gcn5 possesses a HAT domain and a bromodomain and is highly homologous to the C-terminal halves of human PCAF and GCN5L (mammalian GCN5 long form) (60,72–75). Like Drosophila GCN5, mammalian PCAF and GCN5L possess PCAF-specific N-terminal domains (Fig. (Fig.1A)1A) (74). A recent study indicates that GCN5 from the worm Schistosoma mansoni has a similar domain organization (76). Numerous studies indicate that these HATs function as histone- acetylating transcriptional co-activators (4,6,21). Besides histones, PCAF also acetylates non-histone proteins (20,23).
The p300/CBP family is another major group of nuclear HATs that has been extensively characterized (Fig. (Fig.1B)1B) (77–79). Like PCAF, both p300 and CBP are transcriptional co-activators able to acetylate histones and non-histone proteins. Reminiscent of PCAF and GCN5L, p300 and CBP form a pair of homologous HATs in mammals (Fig. (Fig.1).1). Drosophila CBP is larger, but possesses a similar domain organization (Fig. (Fig.1B).1B). As in Drosophila, there is also only one p300/CBP ortholog in Caenorhabditis elegans (78). In Arabidopsis thaliana there are five proteins displaying sequence similarity to the HAT domains of p300 and CBP (Fig. (Fig.1B)1B) (80,81); functions of these novel proteins remain to be determined.
The MYST family of proteins constitutes a third major group of nuclear HATs (Table (Table22 and Fig. Fig.2).2). Compared with the Gcn5/PCAF and p300/CBP groups, the MYST family is larger, more diverse and not so well characterized. Despite their similar HAT domains, MYST proteins play different roles in various cellular processes. In light of their unique structure and function, these acetyltransferases will be discussed in detail in the next section.
In addition to these three major groups of HATs, more than a dozen other proteins have been shown to possess acetyltransferase activity (Table (Table2).2). Like members of the Gcn5/PCAF family, HAT1, Nut1, Elp3, Hpa2/Hpa3, MCM3AP, Eco1 and ARD1 share three or four similar motifs with various N-acetyltransferases and thus belong to the Gcn5-related N-acetyltransferase (GNAT) superfamily (4,6,24,25, 27,82). One of the shared motifs is the classical acetyl-CoA-binding site, which is also present in the MYST family members and ATF2 (6,65,83,84). In contrast, such a motif is absent in other known LATs, so there may be different acetyl-CoA binding modes. Indeed, CDY and CDYL do not have the classical acetyl CoA-binding motif, but display some sequence similarity to several CoA-utilizing enzymes (70).
Most HATs exist as stoichiometric multisubunit complexes in vivo (Table (Table2).2). The complexes are typically more active than their respective catalytic subunits and display distinct substrate specificities (85–88), suggesting that associated subunits regulate the activities of the respective catalytic subunits. In addition, non-catalytic subunits are also involved in recruiting substrates for targeted action to ensure the specificity. Amazingly, one HAT can be the catalytic subunit of multiple complexes. As listed in Table Table2,2, GCN5L forms at least two distinct multisubunit complexes (89–91), and yeast Gcn5 is the catalytic subunit of four complexes (85,92–96; reviewed in 6). Notably, recent studies indicate that Ubp8, a deubiquitinating enzyme present in two Gcn5 complexes, controls the deubiquitination of histone H2B and methylation of histone H3 (97–99). Therefore, the diversity of multisubunit complexes adds another level of complexity to the already diverse superfamily of LATs.
Some known LATs display weak activity towards substrates tested, so an interesting question is whether low levels of activity observed in vitro have any biological significance. One possibility is that the weak activity is not intrinsic, but rather due to an associated HAT. For example, the HAT activity observed with BRCA2 appears to be from associated PCAF (100). It is noteworthy that a weak or null activity observed with a potential LAT in vitro could also be a 'false negative'. A given substrate used may not be the real one since many proteins are acetylated. Moreover, HATs form multisubunit complexes with different activities and substrate specificities (85–87), so activity data obtained with recombinant catalytic subunits could be potentially misleading. Activities of LATs are dynamically regulated by different mechanisms (see below), so the situation could be even more complicated in vivo. For example, neither Sas2 nor its complex acetylates nucleosomal histone H4 in vitro (101), but Sas2 appears to do so in vivo (102,103). Therefore, different experimental approaches are needed to address whether a potential LAT with weak activity in vitro is an authentic one in vivo.
The acronym MYST is from its four founding members: human MOZ (monocytic leukemia zinc finger protein) (83), yeast Ybf2 (renamed Sas3, for something about silencing 3) (84,104), yeast Sas2 (84) and mammalian TIP60 (HIV Tat-interacting 60 kDa protein) (105–107). As illustrated in Figure Figure2A,2A, a third MYST protein in Saccharomyces cerevisiae is Esa1 (essential Sas2-related acetyltransferase 1) (108,109). In Drosophila (Fig. (Fig.2B),2B), there are five members, including Mof (male-absent on the first) (110,111), Enok (Enoki mushroom) (112), Chameau (camel in French) (113) and two uncharacterized MYST proteins (CG6121 and CG1894). In humans (Fig. (Fig.2C),2C), besides MOZ and TIP60, there are hMOF (ortholog of Drosophila Mof), HBO1 (HAT bound to ORC1, a Chameau ortholog) (114) and MORF (MOZ-related factor) (45). Among these, TIP60 is most similar to Drosophila CG6121 and yeast Esa1 (Fig. (Fig.2A–C).2A–C). There are two uncharacterized MYST proteins in A.thaliana (Fig. (Fig.2D)2D) (81). Similar proteins also exist in Schizosaccharomyces pombe and parasitic protozoa (22). Therefore, this family appears to have members in all eukaryotes.
Members of this family possess highly homologous ~370 residue MYST domains (identity, 36–77%; similarity, 54–84%) (Fig. (Fig.2).2). Structural analysis has revealed that the MYST domain of Esa1 uses an acetyl-cysteine intermediate in the acetylation reaction, so the catalytic mechanism involved is different from that shared by members of the GNAT superfamily of acetyltransferases (115). C2HC fingers are present in all MYST domains except for that of Esa1 (Fig. (Fig.2).2). Despite the absence of zinc and three of the four chelating residues, the corresponding region of Esa1 forms a classical TFIIIA-type zinc finger fold (116). The C2HC fingers of Sas3, Mof and MOZ are known to be essential for HAT activity (104,117,118). A mutation in the C2HC finger of Drosophila Enok affects brain development (112). The C2HC finger of Mof binds to nucleosomes in vitro (117), whereas the C2HC finger of TIP60 is essential for interaction with the translocation E26 transforming-specific (ETS) leukemia protein TEL (119). Therefore, C2HC zinc fingers are important in vitro and in vivo.
Besides MYST domains, this family of proteins contains other structural modules (Fig. (Fig.2).2). One such module is the chromodomain (120), conserved among Esa1, Mof and TIP60 (Fig. (Fig.2A–C).2A–C). The chromodomain of Mof binds to roX (RNA on the X) RNAs and targets the complex to the male fly X chromosome for gene dosage compensation (121). In comparison, the chromodomain of HP1 (heterochromatin protein 1) recognizes Lys9-methylated histone H3 (122,123). Therefore, it will be interesting to determine what roles the chromodomains of Esa1 and TIP60 may have. The PHD (plant homeodomain-linked) zinc finger is another recognizable module (Fig. (Fig.2).2). Two PHD fingers are found in the N-terminal parts of Enok, MOZ and MORF (45,83,112,124). The PHD domains of MOZ and MORF are much more similar to each other than to those of Enok and display high sequence homology to Neuro-D4 and Requiem, two potential transcription factors that do not have MYST domains (83). PHD fingers are also known as LAP (leukemia-associated protein) domains and have been found in many chromatin regulators (125,126). Mutations in PHD fingers of several chromatin regulators contribute to a variety of human diseases (127,128). Atypical PHD zinc fingers with similarity to RING fingers have been shown to function as E3 ubiquitin ligases (129), but it is unclear whether this is a common functional feature of all PHD fingers (130). Interestingly, PHD fingers from several chromatin regulators bind to phosphoinositides and are thus implicated in nuclear lipid signaling (131). The PHD zinc fingers of MOZ display phosphoinositide-binding activity, raising the interesting possibility that phosphoinositides act through PHD zinc fingers and regulate the functions of Enok, MOZ and MORF.
Besides PHD fingers, MOZ and MORF share their extreme N-terminal regions with Enok (Fig. (Fig.2B2B and C), referred to as NEMM (N-terminal region in Enok, MOZ or MORF) domains. The NEMM domains of MOZ and MORF, but not that of Enok, display some sequence similarity to the globular domains of linker histones H1 and H5. The functional significance of this similarity remains to be determined. These H1- and H5-like regions, known as H15 domains, may mediate self-association and interaction with core histones and nucleosomes since the globular domains of histones H1 and H5 are known to have similar activities (132,133). In addition, Enok possesses an uncharacterized neurofilament protein-like domain that is missing in other MYST proteins (Fig. (Fig.2B)2B) (112). At the C-terminal ends of MOZ and MORF (Fig. (Fig.2C)2C) are serine- and methionine-rich regions, which are known as SM domains and possess potent transcriptional activation potential (45,134). Therefore, MYST proteins have diverse domain organizations.
Consistent with their diverse domain organizations, MYST proteins exist as distinct multiprotein complexes in vivo. Yeast Esa1, Sas2 and Sas3 are the catalytic subunits of different multiprotein complexes (Table (Table2).2). The Esa1 complex NuA4 contains 12 subunits, including Tra1 (TRRAP homolog 1), actin, Arp4 (actin-related protein 4), Epl1 (Enhancer of Polycomb-like protein 1), Yng2 (homolog of mammalian ING1, for inhibitor of growth 1) and Yaf9 (homolog of the leukemogenic human protein AF9) (94,135,136). Among these subunits, Esa1, Epl1 and Yng2 form a highly active but much smaller core complex, termed Piccolo NuA4 (88). Sas2 and Sas3 are the catalytic subunits of two trimeric complexes: Sas2 associates with Sas4 and Sas5 (AF9 homolog), whereas Sas3 interacts with Taf30 (AF9 homolog) and Yng1 (ING1 homolog) (137–139). Since Esa1 and Sas3 associate with homologs of mammalian ING1, phosphoinositides may bind to the complexes and regulate their functions. In Drosophila, one MYST complex has been characterized (140). Mof in male flies is part of a dosage compensation complex that contains Msl1 (male-specific lethal 1), Msl2, Msl3, Mle (maleless, homolog of mammalian RNA helicase A) and two non-coding RNA molecules, roX1 and roX2. Notably, this is the only HAT complex known to contain RNA. The chromodomains of Mof and Msl3 are able to mediate RNA binding (121). In mammals, the TIP60 complex has been purified and characterized (141,142). It shares similar subunits with the Esa1 complex, including TRRAP (transformation/transcription domain-associated protein), actin, BAF53 (actin-related protein) and EPC (Enhancer of Polycomb-like protein). However, the TIP60 complex also possesses unique subunits such as p400 (SWI2/SNF2-related ATPase), two RuvB-like proteins and DMAP1 (DNMT1-associated protein 1) (141,142). Originally identified as an essential cofactor for the oncogenic transcription factors c-Myc and E2F (143), TRRAP is an ATM/PI-3 kinase-like protein shared by the PCAF (144), GCN5L (89–91), TIP60 (141) and p400 complexes (145) (Table (Table2).2). As the yeast homolog of TRRAP, Tra1 is a common subunit of the Gcn5 and Esa1 complexes (92–96). In contrast, the Sas2, Sas3 and Mof complexes do not possess proteins similar to TRRAP. It is expected that human MOF does not associate with TRRAP, but it remains unclear whether other MYST proteins, such as HBO1, MOZ and MORF, interact with TRRAP.
As suggested by their unique domain organizations and different complex compositions, MYST proteins are involved in regulating various biological processes. In agreement with the fact that the NuA4 subunit Yng2 is homologous to the candidate tumor suppressor ING1 (146,147), Esa1 is important for DNA repair and cell cycle progression (108,109,148). The Esa1 complex has also been linked to epigenetic control, gene regulation and cellular response to spindle stress (135,136). In support of its role in yeast gene silencing (84), Sas2 opposes the action of the deacetylase Sir2 to establish the boundary between euchromatin and heterochromatin (102,103). Sas3 was originally identified as a regulator of gene silencing (84), whereas its complex was recently shown to be involved in regulating transcriptional elongation (137). Drosophila Chameau is important for heterochromatin-mediated gene silencing and Mof is a major player in gene dosage compensation in male flies (110,112). As for mammalian MYST proteins, TIP60 plays an important role in apoptosis and DNA repair (141).
Different lines of evidence suggest that MYST proteins are also targeted to specific promoters to regulate transcription. Through interacting with DNA-binding transcription factors, yeast Esa1 is recruited to regulate the expression of ribosomal proteins (149). The tumor suppressor p53 interacts with ING proteins and its ability to activate transcription in yeast is modulated by Yng1 and Yng2, two ING homologs that are subunits of the Sas3 and Esa1 complexes, respectively (147,150). TIP60 not only functions as a transcriptional co-repressor for TEL (119) and STAT3 (151), but also as a coactivator for androgen receptor (152,153), NF-κB (154) and c-Myc (155). Through a unique NPxY motif within its MYST domain, TIP60 interacts with the WW domain of Fe65 and potentiates transcription mediated by the APP (amyloid-β precursor protein) cytoplasmic domain (154,156–158). Therefore, TIP60 regulates transcription in a context- dependent manner. Originally identified as a protein interacting with ORC1, an ORC (replication origin recognition complex) subunit (114), HBO1 has recently been found to function as a transcriptional co-repressor for androgen receptor (159). Human MOF is part of a complex implicated in activating the B-myb promoter (160). MOZ and MORF possess transcriptional repression and activation domains (45,134,161), suggesting that these two HATs are potential transcriptional co-regulators. Indeed, MORF is present in a transcriptional co-activator complex associated with the nuclear receptor PPARα (162). Both MOZ and MORF physically and functionally interact with Runx1 and Runx2 (161,163,164), two Runt domain transcription factors important for cell growth and differentiation in different tissues (165–168). In agreement with this, down-regulated expression of mouse MORF, known as Querkopf (squarehead in German), leads to defects in osteogenesis and neurogenesis (169). Therefore, compared with members of the Gcn5/PCAF and p300/CBP families, MYST proteins are much more diverse in domain organization, multiprotein complex formation and biological function.
With more information available on the structure and function of different LATs, the regulation of their enzymatic activities has become an important issue in the past few years. Emerging data suggest that multiple mechanisms are involved. First, as an essential cofactor for different acetyltransferases, acetyl-CoA also stabilizes GCN5 and PCAF (42). Second, as described above, formation of stoichiometric multisubunit complexes modulates the specific activities and substrate specificities of different LATs. Third, the enzymatic activities of PCAF, p300 and CBP are regulated by interaction with transcription factors such as p/CIP (170), Twist (171), vIRF (172), Zta (173), HOX proteins (174), PU.1 (175) and the early B-cell factor (176); the ubiquitin ligase MDM2 (177–179); the protein kinase RSK2 (180); and viral proteins like E1A (170,171,181,182), E1B (183), T antigen (184), E7 (185) and Tat (186). Fourth, LATs are subject to covalent modifications such as phosphorylation (65,187), acetylation (37,46), ubiquitination (188–190) and sumoylation (191). Fifth, LATs are degradated by caspases, calpains and ubiquitin-dependent proteasomes (188–190,192). Sixth, subcellular compartmentalization is an important regulatory mechanism for HATs. For example, HAT1 binds to 14-3-3 proteins (193) and TIP60 is sequestered to the cytoplasm in a signal-dependent manner (154,156,158,194). Finally, while p300, CBP, MOZ and MORF possess PHD fingers (Figs (Figs11 and and2),2), yeast Esa1 and Sas3 associate with PHD finger-containing subunits (6,22). PHD fingers are implicated in phosphoinositide binding and may thus provide structural modules for integrating nuclear lipid signals (131), so activities of these acetyltransferases may be regulated by nuclear signaling events.
In addition to the regulatory mechanisms just described, substrate recognition is another point for controlling acetylation. A relevant question is how the specificity is achieved. The classical 'hit-and-run' model can be used to depict how HATs recognize free histone substrates (Fig. (Fig.3A).3A). For example, HAT1 acetylates newly synthesized histones in such a mode (195,196). HATs also adopt 'attract-and-hit' mechanisms. To bring substrates into their physical proximity, HATs and other LATs may either bind directly to their substrates (Fig. (Fig.3B)3B) or interact with their substrates through adaptor proteins (Fig. (Fig.3C).3C). Since physical proximity increases the local substrate concentration, 'attract-and-hit' modes are more efficient than 'hit-and-run' ones. For example, when the WD40-repeat histone-binding protein HAT2 is present, HAT1 is 10 times more active (197). With polymer substrates such as chromatin, region-specific acetylation is sometimes necessary (Fig. (Fig.3D).3D). To specifically modify chromatin, HATs are often recruited to carry out targeted acetylation, which is reminiscent of the indirect 'attract-and-hit' mechanism (Fig. (Fig.3C).3C). In this scenario, subunits associated with a HAT may modulate the targeting specificity. As stated above, incorporation of one HAT into different complexes adds another level of diversity for substrate targeting. Like chromatin, microtubules are polymers, so a tubulin acetyltransferase may also need to be targeted. While the DNA sequence marks the position of a given nucleosome within a chromatin array and thus dictates region-specific recruitment of HATs for acetylation, microtubules do not have such a marking system. Region-specific tubulin acetylation on a microtubule may be achieved during polymerization. As illustrated in Figure Figure1,1, most members of the Gcn5/PCAF and p300/CBP families contain bromodomains, so these acetyltransferases are able to participate in 'relay' reactions (Fig. (Fig.3E),3E), with one enzyme executing the initial acetylation in a polymer substrate to create a binding site for the bromodomain of another enzyme, which then initiates another round of acetylation to trigger a cascade of reactions. This may be one means to execute chromatin domain-, chromosome- and genome-wide acetylation (198). A slightly different scenario is that with acetylating activities coupled to acetyllysine-binding bromodomains, two LATs cooperate with each other to carry out sequential acetylation of different substrates. Such a mode of action has been elegantly demonstrated for p300 and PCAF in their sequential acetylation of Tat and histones (Fig. (Fig.3F)3F) (199,200). Therefore, LATs utilize different means to cope with various types of substrates.
Once a lysine residue is acetylated, a mechanistic question is how such a modification may affect protein function. It appears that both 'loss-of-function' and 'gain-of-function' mechanisms are involved. Regarding the former, acetylation of the ε-amino group of a lysine residue neutralizes the positive charge, so the modification may affect interaction of the lysine residue with DNA, RNA and proteins. Such a mechanism may operate with chromatin since the DNA backbone is negatively charged. Indeed, histone acetylation has been shown to affect the nucleosomal structure and the stability of nucleosomal arrays (201–204). Alternatively, acetylation may render the ε-amino group unable to form hydrogen bonds. For example, Lys11 of Alba is involved in forming a hydrogen bond important for oligomerization, so acetylation of this residue inhibits oligomerization (8,9). Acetyl-CoA synthetase from S.enterica provides another unique example of a 'loss-of-function' effect (10). Lys609 of this enzyme is part of the catalytic center, so acetylation of this residue inhibits the enzymatic activity.
In eukaryotic cells, the ε-amino group of a lysine residue is also subject to methylation and modification by ubiquitin and ubiquitin-like proteins such as NEDD8 and SUMO (Fig. (Fig.4).4). Different modifications are mutually exclusive, thus leading to their potential competition. It has recently been shown for SREBP and Smad7 that acetylation directly competes with ubiquitination for the same lysine residues to increase protein stability (51,205). Acetylation and methylation of histone H3 at Lys9 have completely opposite functional consequences, with the former associated with active chromatin and the latter linked to heterochromatin or inactive chromatin (2,5). Since acetylation blocks methylation, histone deacetylases are needed to remove the acetyl group from Lys9 and clear the way for subsequent methylation and thus the establishment of inactive chromatin (206,207).
As for 'gain-of-function' mechanisms, the addition of an acetyl group to a lysine residue creates a new surface for protein association. Reminiscent of domains that recognize phosphoproteins (208,209), bromodomains function as structural modules specific for acetyllysine-containing motifs (Fig. (Fig.4).4). It has been demonstrated that several chromatin regulators use bromodomains to recognize acetyllysine (210–216). Bromodomains are found in many proteins and sequence variations may dictate their binding specificity (217). Some proteins contain multiple bromodomains, which may cooperate with each other to increase the affinity for binding partners with multiple acetylated lysine residues (211). Acetylation is also known to stimulate the association with proteins that do not contain bromodomains (27,218). Different types of structural modules have been identified for phosphotyrosine (209,219), so an interesting question to be addressed is whether there are additional acetyllysine-recognizing modules (Fig. (Fig.44).
Consistent with the essential roles of LATs in different biological processes, molecular and genetic studies have revealed that these enzymes are also important players in human pathology. Among others, the following lines of evidence strongly suggest that several HATs are directly linked to oncogenesis: (i) the viral oncoproteins such as E1A and large T antigen target p300 and CBP (77,78,220); (ii) E1A also interacts with PCAF and TRRAP, a subunit of multiple HAT complexes (6,143,220,221); (iii) the proto-oncoprotein SYT targets p300 (222); (iv) p300, CBP and PCAF associate with and modify various transcription factors, such as p53, Rb, E2F, H1Fα and E2A, that play key roles in controlling different cellular programs (21,77–79); (v) oncogenic transcription factors c-Myc and E2F bind to TRRAP (143); (vi) the tumor suppressor p53 also binds to ING proteins, homologs of which have been found in HAT complexes (147); (vii) MOZ and MORF interact with Runx1 (161,163,164), the most frequent target of leukemia-associated chromosomal translocations (166,168); (viii) TIP60 associates with the androgen receptor and has been implicated in the development of prostate cancer (152,153,223); (ix) TIP60 is involved in regulating apoptosis and its yeast homolog Esa1 is essential for cell cycle progression (108,109,141).
Among known HATs (Table (Table2),2), p300 and CBP have been considered as tumor suppressors (77–79). Consistent with this notion, monoallelic mutation of the CBP locus is the genetic basis for Rubinstein–Taybi syndrome and patients with this syndrome exhibit an increased risk of developing malignant tumors (224). Biallelic mutations of the p300 locus have been identified in human cancers of epithelial origin (225,226) and exogenous expression of p300 is able to suppress the growth of human carcinoma cells in vitro (227). Moreover, p300 and CBP play distinct but essential roles in hematopoiesis, and mice with inactivated alleles of the p300 and CBP loci develop hematological malignancies (228–231).
Studies of chromosomal abnormalities in leukemia patients have provided additional support for the direct involvement of HATs in human cancer. The p300 and CBP genes are located on chromosomes 16p13 and 22q13, respectively, and have been shown to be rearranged in chromosomal translocations associated with leukemia or treatment-related myelodysplastic syndrome (Fig. (Fig.5A).5A). Known fusion partners are MOZ, MORF and MLL (mixed lineage leukemia) (Fig. (Fig.5).5). The MLL gene, located at 11q23, is fused to the p300 and CBP genes in the translocations t(11;22)q23;q13) and t(11;16)(q23;p13), respectively (Fig. (Fig.5A5A and C) (232–238). These translocations lead to the production of different MLL-p300 and MLL-CBP fusion proteins, in which the bromodomain, HAT domain and Q region of p300 or CBP are linked to the N-terminal part of MLL. The MOZ gene, located at 8p11, is fused to that of CBP in two t(8;16)(p11;p13) translocations (Fig. (Fig.5B),5B), which gives rise to MOZ-CBP proteins containing the N- and C-terminal parts of MOZ and CBP, respectively (83,239). In two t(8;22)(p11;q13) translocations, the MOZ gene is fused to the p300 gene, generating MOZ-p300 fusion proteins (240,241). In the t(8;16) and t(8;22) translocations, the reverse transcripts expressing CBP-MOZ or p300-MOZ are not always produced, suggesting that the MOZ-CBP and MOZ-p300 fusion proteins are responsible for the leukemogenesis. These fusion proteins possess the NEMM, PHD and MYST domains of MOZ, as well as the CH1, KIX, bromodomain, HAT domain and Q region of CBP or p300 (Fig. (Fig.5A5A and B). Notably, the SM domain of MOZ is missing from these fusion proteins.
Besides p300 and CBP, other partners are also involved in the fusion with the MOZ gene. In two slightly different inv(8)(p11q13) chromosomal inversions (Fig. (Fig.5C),5C), the MOZ gene is fused to the TIF2 gene located at 8q13, creating a protein with the N-terminal part of MOZ fused to the C-terminal part of TIF2 (242–244). TIF2 is a member of the p160 family of nuclear receptor co-activators known to interact with p300 and CBP. A recent study indicates that the MOZ gene is also rearranged in a t(2;8)(p23;p11) translocation associated with therapy-related myelodysplastic syndrome, but the translocation partner remains to be identified (245).
The sequence similarity between MOZ and MORF suggests that the MORF gene is rearranged in a manner similar to the MOZ gene (45). Indeed, the MORF gene was recently found to be rearranged and fused to the CBP gene in a recurrent t(10;16)(q22;p13) translocation associated with acute myeloid leukemia (Fig. (Fig.5A5A and C) (246,247). A slightly different t(10;16)(q22;p13) translocation, associated with therapy-related myelodysplastic syndrome, also leads to fusion of the MORF gene to the CBP gene (247). The resulting MORF-CBP fusion proteins are structurally similar to the MOZ-CBP and MOZ-p300 fusion proteins described above.
All of the above chromosomal abnormalities suggest that aberrant acetylation by mistargeted HATs plays a causative role in leukemogenesis. Indeed, one MLL-CBP fusion protein has been analyzed in mice and found to generate a myelodysplastic syndrome that evolves into myeloid leukemia (248). The bromodomain and HAT domain of CBP are the only modules needed for the leukemogenic activity, suggesting that the N-terminal part of MLL directs the two CBP domains for aberrant acetylation and subsequently leads to leukemogenesis. MLL is processed by Taspase 1 cleavage and the resulting N- and C-terminal fragments remain associated within the same histone methyltransferase complex (Fig. (Fig.5C)5C) (249). It is unclear whether the MLL-CBP fusion proteins are still able to bind the processed C-terminal fragment of MLL. Potential targets of MLL-CBP are members of the HOX gene family (250,251), so MLL-CBP may lead to abnormal expression of these genes and cause the subsequent development of leukemia (Fig. (Fig.66A).
A MOZ-CBP fusion protein from t(11;16)(q23;p13) has been characterized (161). This fusion protein inhibits Runx1-dependent transcription and blocks the differentiation of murine myeloid M1 cells to macrophages. Moreover, the HAT domain of CBP appears to be important for the repressive activities. Two slightly different MOZ-TIF2 fusion proteins from inv(8)(p11q13) chromosomal inversions have recently been shown to display oncogenic potential in both in vitro transformation and in vivo transplant assays (118). The C2HC zinc finger of MOZ (Fig. (Fig.2)2) is essential, whereas its acetyl-CoA-binding motif only modulates the penetrance and phenotype of the resulting diseases. For TIF2, only its CBP-interacting domain is essential, so the recruitment of CBP to MOZ is responsible for leukemogenesis. This finding suggests that the underlying molecular mechanism is very similar to that used by MOZ-CBP. Therefore, both MOZ-CBP and MOZ-TIF2 may repress Runx1-dependent gene expression and cause the development of leukemia (Fig. (Fig.6B).6B). Although both MLL-CBP and MOZ-CBP are leukemogenic, their mechanisms of action are quite different, with the former being an activator and the latter functioning as a repressor (Fig. (Fig.66).
As a common post-translational modification, lysine acetylation is known to occur not only at the N-terminal tails of core histones but also within other eukaryotic proteins, including about 40 transcription factors, a chromatin remodeler, one DNA replication factor, three DNA metabolic enzymes, a signaling regulator and a cytoskeletal protein (Table (Table1).1). In addition, viral proteins are also Nε-acetylated. Most of these proteins are nuclear, so it will be interesting to examine whether lysine acetylation plays a wider role in the cytoplasm. Intriguingly, this modification has been found in at least three bacterial proteins (Table (Table1),1), so it is not just unique to eukaryotic and viral proteins. Consistent with the wide spectrum of substrates, a highly diverse superfamily of LATs has been identified (Table (Table2).2). However, enzymes responsible for the acetylation of α-tubulin, Alba and acetyl-CoA synthetase remain elusive. Most known acetyltransferases are catalytic subunits of multiprotein complexes. With distinct sets of subunits, one acetyltransferase can be the catalytic subunit of different complexes, thereby adding another level of diversity. Biochemical and molecular approaches have been and will continue to be fruitful in analyzing the function and regulation of acetyltransferase complexes (6). To understand how lysine acetylation regulates different cellular processes in vivo, genetic analysis has been invaluable to distinguish between functional differences of homologous HATs. Gene targeting in mice has revealed that GCN5L but not PCAF is essential in early embryonic development and that p300 and CBP play distinct roles in hematopoeisis (228,230,252,253). Therefore, such an approach shall continue to yield novel insights into the biological function as well as the spatial and temporal regulation of these and other LATs.
As for molecular mechanisms by which lysine acetylation exerts its effects, one major advance made in the past few years is the discovery that bromodomains are able to recognize acetylated lysine residues (210–216). Many proteins possess such modules (254,255), so one interesting question is whether all bromodomains have such an ability. If so, one major challenge would be to determine the binding specificity of different bromodomains. Degenerate peptide libraries have been successfully used to determine the binding specificity of structural modules involved in cytoplasmic signaling (256). Similar strategies should be valuable for establishing the binding specificity of different chromodomains and understanding their functions in recognizing specific acetylation signals.
The genes for CBP, p300, MOZ and MORF are rearranged by leukemia-associated chromosomal abnormalities (Fig. (Fig.5).5). Some of the resulting fusion proteins have been shown to be leukemogenic (118,161,248). TIP60 has been implicated in the development of prostate cancer (223) and it also interacts with and modulates the function of the cytoplasmic domain of APP, an important regulator of Alzheimer's disease (154,156–158). Expanded polyglutamine tracks, a root cause of Huntington and other polyglutamine diseases, target the acetyltransferase activity of CBP (257–260). In addition, lysine acetylation affects the important roles that various proteins play in other human diseases (Tables (Tables11 and and2).2). For example, acetylation of HIV Tat is an essential regulatory step for virus production. Therefore, LATs play important roles in the pathogenesis of leukemia (Figs (Figs55 and and6)6) and other diseases.
Small molecule inhibitors and activators of histone deacetylases have been extensively explored for the treatment of cancer and other human diseases (17,261,262). The direct involvement of LATs in leukemia and other diseases suggests that small molecules able to modulate their enzymatic activities should be of therapeutic potential. In contrast to histone deacetylase inhibitors, only a few small molecules have been shown to modulate the activity of HATs (263–265). Model organisms such as S.cerevisiae possess various HAT orthologs (Figs (Figs11 and and2)2) and may be used to screen for cell-permeable compounds using similar assays to those described for histone deacetylases (266,267). The resulting compounds will yield lead structures for further optimization with iterative rounds of structure-based molecular design and chemical synthesis. Therefore, studies of the fundamental process of lysine acetylation will not only yield important further insights into how post-translational modifications regulate various cellular processes, but also shed new light on the development of novel therapeutic means for the treatment of leukemia and other human diseases.
This work was supported by funds from the Canadian Cancer Society through the National Cancer Institute of Canada.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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Tanja Kolbe and Paul Boll at the German junior championships 2006 in Berlin at the original dance


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AN:Hello, all! Welcome to my new chaptered story! It's all done already, all I have to do is upload it, which I will do by uploading a new chapter every Friday.
This was done as a sort of a collaboration between myself and the wonderful Maravelous. I told her I was planning on a 1920s fic, and she came up with an awesome plot and sent me basic outlines for each of the ten chapters.
Oh, by the way, it's Maravelous's birthday, so go wish her a happy one! If you've never heard of her (you poor, poor depraved reader), go check out her stuff. She's an amazingwriter. :D
Well. Without further ado, may I present you with:
Strawberry Daiquiri
Chapter One; Tastes that Linger
Ten Years Ago...
"Dear, hurry up!" a woman urged as the child behind her tripped over her feet, striving to reach her mother. The woman extended her hand out, and the little girl latched on to it with strength surprising for a five year old.
She turned her chubby-cheeked face up to the elegant woman whose hand she held. "Thanks for taking me shopping with you, momma!" the girl giggled, pressing close to her mother's side.
The woman smiled down at her and ruffled the chocolate brown hair that flowed freely to her daughter's shoulders. "Anytime, Tenten," she promised. Maybe it wasn't so easy to get around with a young girl latched onto her, and perhaps her shopping trip would take a little longer than usual, but those were a small price to pay for her dear daughter's smile.
"Spare change, please?" The request caught Tenten off-guard, and she tripped over her own feet. A firm hand that wasn't her mother's steadied her. "Careful there, little lady," the homeless man said cheerfully, his wide grin revealing a mouth that was more full of gums than of teeth. Tenten's eyes drifted down towards the crowd walking past. No one was stopping, despite the man's soft requests. The little girl sighed, a frown falling across her features. Not a single person was willing to even pause?
"Here, this is all the change I have, but I hope it helps," the melodic voice of Tenten's mother murmured, breaking through the girl's thoughts. A slender hand pressed a bill and some coins into the man's dirty hand, and an even broader smile broke across his face.
He carefully placed the money in a pocket of his worn clothing. "Bless you, lady, and your little daughter, too. I hope she grows up kind like her mother," he said, patting Tenten on the head. The girl was pulled after her mother, waving over her shoulder at the homeless man.
"Momma, no one else was stopping. Why did you do that?" the girl asked, a curious look etched across her features.
The woman gently pulled her daughter closer to her side. "What everyone else does shouldn't decide what I do. Besides, I had the change to spare, and he needed it. Is that not reason enough?" she asked mildly.
Satisfied with her mother's reasoning, Tenten nodded. Yet it wasn't long before she picked up her face again, her brow furrowed. "Didn't it make you sad to see him like that?" she wondered.
"A little. But maybe he'll be happier in the future. That's the kind of future I'd like to make possible for him."
Present Day...
Tenten frowned as she snapped the handcuffs on the rather dirty wrists of the man in front of her. Honestly, just because she was one of the few female police officers didn't mean she was any less capable that any of the males, she thought angrily as she yanked the man to his feet. And she certainly didn't do anything to deserve this lack of respect, either.
"You're under arrest for loitering and possession of alcohol. Or did being homeless keep you from understanding that prohibition went into effect quite a few months ago?" she asked, her tongue sharp in her anger.
The man at least wasn't resisting, she though, almost sighing in relief. All she would have needed was for someone to resist arrest for some of her male co-workers to exhort that she needed protection. Hell, she had earned her badge just as much as all of the rest of them. And she was a better officer than some of them. Stupid, misogynist, chauvinist pigs, the lot of them... well, perhaps not all, but still. "Come on, into the car, and don't even try anything idiotic."
He allowed himself to be pushed into the backseat, and Tenten slid into the driver's side of the Ford model-T. "What was that? Speak up," she demanded. All she'd heard was some incoherent mumbling from the back. Odd. He hadn't seemed drunk.
"I said 'what happened to compassion?'" the man replied steadily.
Tenten tried to keep from snorting. A rather crooked smirk appeared on her lips, and she started the automobile. "This is real life, not a drunken fairy tale. Now, where'd you get the bootleg from?"
. . .
Neji almost growled as he found himself shoved rather brusquely through the double doors of Ichiraku's. "Honestly, Temari, you're constantly dragging Miss Hinata and me here. It's becoming a little bit tiring, and more than slightly annoying," he scoffed.
The blonde in question just laughed and waved at the waiter near the front doors, a man named Kiba whose job it was to seat those not familiar with the restaurant. "Really, Neji. Come on, it's not like you don't like it!" she retorted cheerfully. Neji smiled in spite of himself as Hinata just giggled and pushed a lock of dark hair out of her eyes. Meanwhile, Temari's sharp eyes lit upon a table occupied by only two others.
She swooped down on it, pulling Neji and Hinata along with more force than one would suspect from the young woman. They all took seats, facing the stage in front of them. A jazz band blasted a sultry tune into their ears, but the microphone stand at the edge of the platform was unmanned and looked strangely lonely. "It certainly took the three of you long enough," a dark-haired man drawled, his cigarette tipping precariously from his fingers.
Temari scowled rather fiercely at him. "Well, sorry. But you weren't the one stuck with bringing the Hyuugas here. No offense, Hinata, I was alluding more to your cousin over here," the blonde woman added quickly, lest the sweet Hinata worry that she had caused Temari any undue troubles. As Neji glanced to his left, he surmised Temari shouldn't have bothered.
His cousin was glancing sweetly and surreptitiously over at the blonde man on her left, and a light blush was spread across her cheeks and the bridge of her nose. "Um, hello, Naruto," she said softly.
"Hey, Hinata! How are you?" he replied, his excited and slightly grating voice a marked contrast to her demure tones. Vaguely, Neji wondered if he should castrate the boy now and save himself some time. Then again, Hinata might get upset. But if it was for her own good...
Neji's attention was shifted from that fascinating topic when he noticed that Ichiraku, the owner of the bar, stood over the table, grinning. "Will you all be having the usual?" the man asked, looking over the group.
General murmurs of assent were his reply, but Neji frowned slightly and then shrugged. "No, I think I'd prefer to try something new today," he replied.
"How about I surprise you?" Ichiraku asked after a moment of thought. Neji simply nodded his head. Why not? "Well then," the owner continued, "I'll be back with your drinks in a moment." Just then the band finished their song, and the silence was greeted by rather unenthusiastic applause. A small frown appeared on Ichiraku's face. Clearly the diners were looking for something more than mere instrumentals, Neji thought, slightly amused. As he turned to go, the man nudged Shikamaru and then ambled off away from the table.
Shikamaru heaved a great sigh and took a drag out of his cigarette. "No way. I'm tired today, and I want to get a chance to finish my ciggy. Besides, we just ordered our drinks. They won't be out for a few more minutes," he hedged.
Whirling to face him, Temari slapped Shikamaru across the top of the arm harshly. "Stop whining and get your ass up on stage. Honestly, you can't even act like a man," she complained, her teal eyes narrowed.
"Fine, fine," Shikamaru sighed as he stood. "Naruto, are you going to help out?" he asked as he grabbed his fedora from the table and placed it on his head.
Naruto literally bounced out of his seat, nearly upsetting the table in the process. "Yeah, Shikamaru!" he said, his voice excited. Hinata's quiet giggle was only noticed by Neji, who rolled his eyes at the antics of the blond. "Let's go then!" he said, vaulting up onto the stairs, followed by Shikamaru, who ascended the platform in a exponentially more calm manner.
Both of the men were seen by the visitors at the restaurant. Neji noted that the newcomers simply sat and stared, but those who were familiar with the two clapped and cheered as Shikamaru approached the lonely microphone and Naruto opened the case for his trombone.
Turning to the band, Shikamaru tilted the brim of his hat slightly over his eyes. "So, do you know 'I Can't Give You Anything But Love, Baby'?" he asked cooly. Neji smirked as he heard Temari huff and mutter something unintelligible. A cacophony of agreement reached his ears, and Shikamaru smiled lazily and moseyed over to the microphone. "Naruto'll lead you guys," he told them over his shoulder. "Sorry about that part," he muttered into the microphone. Naruto whined some inane reply and Neji heard the crowd buzz with amusement around him.
For the first few bars, Shikamaru's smooth voice rolled like velvet over the entire restaurant. Good, as always, Neji though, a small smile starting to tug at the corners of his lips. Under the table, his foot was tapping in time to the music, just as Naruto started to blast clear notes through his trombone and the rest of the band slowly joined.
Even more interesting than the wonderful music, in Neji's opinion, was the starry-eyed looks playing across the faces of the women in Ichiraku's. Even Temari and Hinata, try as they may to hide it, seemed a little dazed and more than slightly captivated. As Shikamaru's gaze caught Temari's, Neji almost laughed. Temari, one of the more independent-minded women he ever knew, was blushing, the pink color splashed across her cheeks and the bridge of her nose.
Speaking of pink, a rather fruity-smelling, pink-red drink found its way in front of Neji. The rest of the drinks around the table, even Hinata's, looked somber and businesslike in comparison. "Interesting drink, Neji," Temari snorted, her attention drawn enough by his cocktail for her to leave Shikamaru's voice behind for a moment. "Kind of a dame's drink, isn't it?"
Ichiraku, who had finished passing around the drinks, shot the blonde woman a look. "It's alcohol; how is it feminine?" he demanded.
Temari, who had no answer for that question, merely shrugged her shoulders. "Seeing as I paid for it, I see no reason not to drink it," Neji retorted, practical as always. Still, he was a bit apprehensive as he raised the cocktail to his lips and took a sip of it. He found the taste to be quite pleasing, fruity but slightly acidic and with a slight burn. Ichiraku paused to tell him it was a new drink called a Strawberry Daiquiri before turning back to chat with Hinata and Temari.
As Hinata giggled and Temari chuckled, Neji looked around the bar. Already he was slightly bored. Although Ichiraku made excellent drinks and Naruto and Shikamaru were amazing musicians, Neji still found merely sitting around and listening to others talk about inane things to be a waste of his time.
The opening of the main doors, then, was quite a welcome distraction.
There were some tastes that lingered in Neji's mind; the taste of his first kiss, the sour taste of his uncle's criticisms, the taste of smoke in his mouth the first time he tried a cigarette. The taste of the strawberry daiquiri and something else, something unidentifiable, swirled in his mouth when he saw her for the first time, and he knew that the taste would be one of those that would linger on while others faded.
The sun was setting behind her, giving a strange sort of glow to her while making it hard to stare too long, making his eyes hurt when he tried to drink her in. While the police uniform wasn't something he'd quite wanted to see on such an adorable woman, it somehow only served to add to her charm, like the dark brown hair pulled into a bun at the back of her neck, or the large brown eyes that narrowed as she seemed to ask something of Kiba.
Neji's eyes were fixated on Kiba's finger as the younger man slowly pointed to Ichiraku, who was still too engrossed in conversation to notice what was going on. Neji even continued to stare unabashedly as the policewoman fixed her hands on her hips and made her way towards his table.
. . .
Well, if the homeless man was expecting to get alcohol from this place again, Tenten thought, he was in for a rude awakening. She was about to crush this tiny establishment.
Just as she reached the table where the owner was, the band finished their song, and Tenten watched as a man in a fedora slouched off the stage while his blonde friend literally vaulted off of the platform and to the table she was at. "Are you the owner of the bar?" she asked Ichiraku, tilting her chin slightly up so she could see directly into his eyes.
"Yes, I am," Ichiraku replied, his voice not a bit hesitant. "Is there any problem here, officer?" he asked, the last word spoken with a sarcastic lilt that wasn't missed by the policewoman.
Tenten tried to keep from rolling her eyes. "Other than this being a gin mill, no. Did you know that there was a homeless man I arrested today who told me–"
She was cut off by the blonde man, who appeared to have just keyed into the fact that she was even there. "Doll, how about we switch roles and I'll handcuff(1) you," he suggested, his eyebrows wiggling in a way that reminded Tenten slightly of her chief and one of the other boys at the police station.
"Beat it, drugstore cowboy(2)" she replied, her voice dry. He pouted, and a dark-haired, well dressed man gave a quiet chuckle. He was quite handsome, she noticed, but her face remained stern as always. "Something funny?" she asked.
The man shrugged, all elegant ease. "Yes, it is, at least a little bit. Anyway, how do you know Ichiraku isn't simply serving soda? Or there are virgin forms of most drinks; it could be one of those."
Tenten eyed the man. "Well, if you're so confident that you're not breaking prohibition, let me smell your drink," she demanded, detaching a hand from her hip to hold out as she waited for him to give her the alcohol. The man wasn't as stupid as she had expected for him to be, being in a speakeasy. He passed her the pinkish beverage without anything more than a sigh. Tenten took a quick whiff of the drink; it definitely was alcoholic. She whirled to the owner of the cheap, dirty little bar, ready to accuse him.
"Look, officer," the man who'd been singing interjected, his voice calm and sounding tired, "give him a break. It's not quite like anyone here is ossified(3). Besides, it's a small bar; what harm could it possibly do?" he reasoned cooly.
Brown eyes narrowed and flickered to the jazz singer. "I already had one taken in today for bootleg that he bought here. Besides, I'm one of the first female police officers. Everyone's watching me, looking for me to slip up just once so they can say that dames can't do this job. I will not have any drunken accidents in this town on my watch," she snapped as she half-slammed the strawberry daiquiri onto the table.
"What a wet blanket," a dirty blonde woman muttered. "Anyway, didn't your mother teach you any compassion?" she asked.
"Well, my mother's dead. What's your excuse; did yours not teach you to think before you spoke?" Tenten retorted sharply.
The other woman only smiled bitterly. "I manage to think before I speak just fine; mine's dead, too, after all."
Before Tenten had a chance to reply to that, the man in a fedora laid a hand on her shoulder. "That aside, it's not like Ichiraku openly hands out hooch to anyone who comes in the doors. People have to ask him, and they have to be people he knows."
She didn't even bother to keep from rolling her eyes this time. "Prohibition doesn't exclude people who know the owners of speakeasies."
"How is alcohol even evil, like you people try to make it out to be?" he demanded with a sigh.
"Well, aside from impairing judgement, doing bodily damage, and making people act even more dumb that they usually manage to, I suppose it's not evil at all," she retorted. Tenten could feel her temper rising as this man tried to convince her that alcohol wasn't bad. What next, would he try to convince her it would be helpful? She almost snorted at the very thought.
Sighing, the man pulled off his hat and held it loosely in his hands. "If people don't get alcohol from here, they'll go to another joint and get it from there. You can't prevent people all around the world from getting their booze."
Tenten gritted her teeth; surely if he was smart enough to put forth these arguments he was smart enough to know not to bait a police officer. "Listen here. Maybe I can't enforce prohibition all around the world. However, I can and I will shut down this little gin mill and all the rest of them in this town," she snarled.
The man who had been trying to convince her otherwise sighed and placed his fedora back on his head, adjusting it slightly as he appeared to think. "Listen," he sighed, "I didn't mean to anger you. You should give us some time to prove that Ichiraku's won't cause any 'drunken accidents'. Three weeks, and then you can make your decision."
As she drummed her fingers on her hips, Tenten thought the proposition over. She'd been a little bit bored lately, and it was hard to find some decent company. The man had already proved himself to be levelheaded and at least passably intelligent, and the others couldn't be too bad. Besides, she could make any decision at the end of three weeks. He wasn't telling her she had to make a choice either way.
"I suppose I agree," she said after a moment, tilting her chin up even as she met the singer's dark eyes. "I'm not making any promises either way, though."
"Swell," the man said, a slow smile breaking across his face. "My name's Shikamaru, by the way. This is Neji, Naruto, Hinata, Temari, and Ichiraku, of course," he said, gesturing to each person in turn. "How about you come tomorrow to listen to me sing?" he invited.
It took her only a moment to consider. "Fine. I need to keep an eye on this place, anyway, if I'm to honor this deal," she said. Then she turned to go. She'd spent too much time in this little joint; there were other places to go, other criminals wandering around somewhere, no doubt, she thought.
A low voice cut into her thoughts. "Why are you even bothering us?" the man Shikamaru had introduced as Neji asked evenly. "Being a female police officer is ludicrous enough. Why do you find the need to dabble in other people's lives, as well?" he asked. The girl named Hinata tried to hush him, but he'd already said his piece.
"Listen here," she cut in, her voice cold. "Woman or not, I am still a police officer and I still have the power to arrest you on the spot. It's a power which I will certainly not hesitate to use if you have the poor judgement to insult myself or even my gender again. Are we clear, Neji?" she demanded, almost amused as he tried to match her glare for glare. She was the one with the handcuffs. The pale-eyed man shrugged and muttered something that she didn't even bother trying to listen to.
Without another word, she turned on her heel and stalked out of Ichiraku's. The taste of the place, the ambiance and the people, remained on her tongue, a feeling she couldn't get rid of. And even as that taste lingered, the smell of the strawberry daiquiri permeated her nose, and she couldn't help but wonder if it tasted as good as it looked.
Tenten shook her head as she close the door behind her, just barely catching the wailing of a trumpet flowing through the crack in the doors before she firmly shut them. It was silly to wish for a drink of the liquor, of course. Especially when she had her own reasons for trying to be a police officer and for enforcing prohibition.
She wanted to keep the place clean and safe for the residents, even if they weren't pleased about her so-called intrusion. She had to do her best for her town. All for the sake of her mother. Tenten pulled the door to her car open and started the automobile.
She had dreams of her own. And a low-class bar wasn't going to ruin them.
. . .
1920s slang dictionary!
1) A double entendre. Handcuffs in the literal sense, but also in 1920s slang, in which it means an engagement ring.
2) A guy who hangs around street corners trying to pick up girls. Desperate.
3) Drunk.


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KNIGHTS Coolangatta, Australia
Formed in 2010 Knights Like These from Coolangatta (Gold Coast) Australia strive to be evermore honest and to the point with every release. With integrity and and self growth being their main motivators, it has set them on a path of perpetual enrichment from their craft. Their only goal is for you to share it with them. ... more
Streaming and Download help


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Rdio Opens Service to Developers with Bounty Incentives
By Antony Bruno
Cloud music subscription service Rdio is opening up its service to developers so that they can add on-demand streaming music capabilities to their apps.
The open API program is good for Web apps and mobile apps that can now add the ability to search and stream all songs and artists in the Rdio catalog. And Rdio is offering a bounty to participating developers for every new Rdio subscriber that joins as a result of discovering the service through any app it's integrated with.
This is an important feature because without an Rdio subscription, users accessing the on-demand streaming feature in any such apps will only be limited to 30-second song samples. Participating developers also get a cut of any a la carte download sales.
Early users of the Rdio developer program include AOL, for its newly announced Play app, Grab.com, Tweetlouder, and the Grammy MusicIsLifeIsMusic website


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We have been approached by a NZ librarian who is visiting the UK from Te Totara Primary School- she says "My name is Michelle Simms and I'll be visiting London next year as part of a scholarship I have received. I'm a primary school librarian in Hamilton, New Zealand and I'm very interested in the work that is being done in UK school libraries around reading for pleasure. As part of my trip I would love to visit some primary school libraries.
I am going to be based in London, but I will also be happy to visit other places as long as they are near public transport. I will be arriving on 27 February and leaving on 17 March."
If anyone can help Michelle by offering a visit then please contact her direct - firstname.lastname@example.com
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Free shipping
All orders over $69
All prices are in All prices are in USD
Laredo Women's Shortie Sapphyre Cowgirl Boots
Laredo reaches far on the style spectrum creating western footwear for the classic or modern cowgirl. The Laredo Sapphrye boot is just plain adorable and feminine for her. A vivid blue leather is highlighted with a modest stitch design. Shorter 6" shaft adds popular flair. Comfortable cushioned insoles. Finished with a medium cowboy heel.


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John Anders Johnson
John Anders Johnson was a member of the Wisconsin State Assembly and the Wisconsin State Senate.[1]
Johnson was born in Telemark, Norway on April 15, 1832. In 1852, he moved with his family to Pleasant Springs, Wisconsin. He married Karen Kristie Thompson in 1856. They would have a daughter before her death in 1860. On October 31, 1861, Johnson married Kaia Nicoline Marie Kildahl. They would have six children. He died of a stomach ulcer on November 10, 1901 and was buried at Forest Hill Cemetery.
Johnson was a member of the Assembly in 1857. He served as Clerk of Dane County, Wisconsin from 1861 to 1869. From 1873 to 1874, he was a member of the Senate. Other positions Johnson held include Town Clerk and Town Chairman of Pleasant Springs and justice of the peace.
Outside politics Johnson owned a company that produced farm machinery, having previously been a farmer.
- ^ A. J. Turner, ed. (1874). The Legislative Manual of the State of Wisconsin (13th ed.). Madison, Wis.: Atwood & Culver. p. 449. Retrieved 2015-06-17.
External links[edit]
- Politicians from Telemark
- Norwegian emigrants to the United States
- People from Dane County, Wisconsin
- Businesspeople from Wisconsin
- Farmers from Wisconsin
- Wisconsin State Senators
- Members of the Wisconsin State Assembly
- Mayors of places in Wisconsin
- City and town clerks
- County clerks in Wisconsin
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My name is Phill and I started flying with Monarch in 2005. I flew from MAN for 5 years and have been in BHX since 2010. As Cabin Manager I help to motivate a team of cabin crew to achieve the daily business goals; to achieve 100% customer satisfaction, work towards the OTP (On Time Performance) goals and to mentor crew members to be the best they can be.
As I fly with different crew each day, no day is similar I getting to meet new crew whether that is crew or passengers. Working my way through the ranks to Cabin Manager and with 10 years' experience I believe I have learnt many new skills and approaches in how to read people and therefore understand the needs of our customers. Getting to know our customers in a short flight is easy with a smile and a genuine "hello"!
Monarch isn't always like going work; it's going for a nice day out with new friends. There is very much a family feel to Monarch. My top tip for anyone wanting to work for Monarch is to be yourself. We all have our own individuality and it's only when we bring these all together that we create the Monarch experience.
Working with Monarch for 10 years I have had the chance to expand my skills through in house training; I have taken courses that select new recruits, help coach crew, assist with training, become a VIP flyer and assisting on promotion days to advertise the Monarch brand.
Monarch is very much a family feel airline and so understands the needs of passengers. As a brand we understand the journey that our customers take isn't just the flight itself but it's the whole adventure from booking the flights, the checking in, the flight and the holiday itself. It's by bringing the whole trip together as one; delivering an exceptional service.


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A Chilly Autumn With Cool Fun
You know it's autumn time when the wind starts to blow, and orange leaves are falling down from the trees, and your cute cat is purring at your lap. If you also have a free afternoon it is even better – you can seat on your porch, relax with the gentle breeze, and have some fun activities that would also be beneficial for you.
For example, one cool and calming activity would be doing some coloring for adults pages. The coloring pages are photos in all shapes and styles that you paint, and get a beautiful art work that you've made yourself! Is addition, this activity is very calming and you can enjoy it's meditative effect before doing something more exciting – like entering your favorite online casino .
Another cool activity you can enjoy from is solving some great mazes . The mazes are all over the internet, and they are a challenging, great puzzles. You can find any maze you like, and start solving it.
After doing some of the activities such as coloring for adults or maze solving, you might as well enter your favorite online casino games and start playing! You should try as much as them as possible before entering the casino games and see the effect for yourself!
Playing And More
Playing at the online casino is a great experience. Your free time can be spent with enjoyment and fun, as well as earning some money! Most of the classic casino games can be found in almost every online casino , so you can find your favorite game.
But before entering the online casino , you can do some things to improve your winning odds. For example – you can do some coloring for grown ups pages. This meditative activity, just like petting your cat or cats , can be very calming and enjoyable.
The next cool activity you can do before playing would be maze solving. The mazes are extremely common and popular, cause they are both fun and challenging. Solving a maze can also improve your concentration and help you with earning more.
So next time you enter your favorite online casino , you should try to enter the coloring for adults pages, find your favorite coloring pages and try them, or you can solve some mazes and see how it improves your skills. If you try it a couple of times before start playing at the online casino games you might find out that you even gain you more money!


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Sterling 72245100-0 - 72245100-0 ACCORD END WALL
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Floor Coverings International is your local solution for solid hardwood flooring in the greater South Denver area. Solid hardwood is differentiated from engineered hardwood because it is crafted from a single piece of timber. This makes solid wood highly durable and allows it to be sanded and refinished multiple times without losing quality.
Our team of flooring specialists will be there for you from start to finish from the product selection, design, and installation of your new solid hardwood flooring. We have a wide range of experience in designing and installing many different brands, species, and styles of hardwood floors.
Advantages & Disadvantages
One of the main advantages of solid hardwood flooring is that it is easy to repair, since it is all made from the same piece of wood. Also, solid hardwood will age naturally and beautifully as the years go by in your home.
On the other hand, solid wood flooring is very susceptible to changes in humidity, so it shouldn't be installed in any basement-level rooms. If you are trying to install a hardwood floor in your basement, we would suggest engineered hardwood flooring. Most installers compensate for solid hardwood's ability to expand and contract with humidity by leaving a small gap between the flooring and the wall.
Solid wood comes in a huge selection of species, from traditional oaks and maples to more exotic African and South American options. Because we have partnerships with leading flooring manufacturers, we have access to all the top solid wood brands and products.


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In case you couldn't tell from the general tone of my food posts, I really like mozzarella cheese. I mean, I really, really like mozzarella cheese. Fresh mozzarella is soft, almost spongy, mild. Smoked mozzarella is firm, dense and rich. Melted mozzarella is springy, tender and chewy in the best way. I could go on but by now the cheese people are drooling!
Earlier this week (Tuesday?) I was at our local market and spotted the display of mozzarella. I looked around quickly for lettuce, tomatoes and a short line. Yes, express checkout! I called Lesli, gave her the goods and went off to run errands. When I got back she had made one salad for herself using fresh tomatoes, and one for me using sun dried tomatoes. My sun dried tomatoes came from a jar, with olive oil. Before I took my first bite, there was a drizzle of fancy oil – a hostess gift from Chela, when she came to visit us in New Jersey a few months ago.
Wednesday, I went out for prosciutto. Lesli got inspired and made salads again. This time she used prosciutto in mine, and still kept hers the same.
If you want precise measurements, you're about to be disappointed. You should know by now, in our house whatever looks good ends up on the plate. Recipes are suggestions.
So here's what Lesli did for her salad – which was almost a traditional Caprese salad. For truly traditional Caprese salad, check out what Pioneer Woman did – it's an amazingly beautiful salad, with ingredients that are easy to find.
Lesli used
Romaine lettuce (chopped)
Fresh juicy tomatoes (sliced and chopped into half-inch cubes)
Mozzarella cheese (sliced and chopped into quarter-inch cubes)
Rinse and dry lettuce. Top with tomato and mozzarella. Drizzle with olive oil, balsamic vinegar, oregano, red pepper flakes – or if you're lucky, red-pepper-infused oil.
What else can a girl want? Friends to share it. Or maybe a second plate. I have to properly thank Lesli for making that second salad the next night, with prosciutto! The salty tang and texture of prosciutto make it another favorite of mine. But that's a story for some other time. Maybe next week?
This post is part of the Sabroso Saturday blog hop originated by MsLatina. Check in at her site for links to recipes from other bloggers across the country, and from all corners of the internet. There's no telling what everyone will cook!


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For the confident, daring & unique bride who is not afraid to shine on her special day. The Angelina encompasses all the right elements of a sexy wedding gown. It is made exclusively by us with soft stretchy intricate European floral lace and features a seductive open low back with adjustable lace-up detail. It's beautiful design with a low v-neckline that highlights the décolletage and open back that concludes just above the hips with delicate lace trimmings is complimented by a gorgeous 0.8 metre long lace trimmed train.
This affordable yet luxe gown was designed to highlight the unique shape of each body, with soft fabrics that hug the natural curves in all the right places in the most flattering way. Designed as a gown to be worn by the modern bride, the sexy bride, the untamed bride, the boho bride or the effortless bride.
Fully lined with a double layer of soft matte jersey fabric (instead of your usual satin fabric without stretch that's unbreathable, sticks & bunches up) with soft floral lace over the top. This dress has the perfect amount of stretch to allow for comfortable wear all day and night long. You'll never want to take it off. Truly for any bride who isn't afraid to own their style and shine at their wedding just like a graceful Goddess.
Our exclusively sourced European floral lace is available in Ivory. Stretch lining is available in Ivory, or for those who wish to have an understated romantic hue, Champagne, Nude or Blush lining is also available.
Note: We fully understand the jitters that come with choosing a bridal gown, which is why we try to make it as easy & stress-free process in ordering your dream gown & offer our full support to you throughout the ordering process . We are available to chat at any time via e-mail, phone or Skype and would be thrilled to offer styling advice or assist with any inquiries you may have. Visit our contact page to get in touch with us.
Style Details:
- Soft intricate European floral lace
- Gorgeous eyelash lace trim edging along front v-neckline & low back
- Fully double-lined with soft stretch jersey fabric
- Sensuous v-neckline to highlight the décolletage
- Plunging low open back with adjustable lace-up straps
- Built in bra cups for support; A, B, C, or D
- Stunning mermaid style 0.8m long lace edging train
- Optional center slit


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Svaman at Wikia
Welcome to the Svaman mini wiki at Scratchpad!
You can use the box below to create new pages for this mini-wiki. Make sure you type
[[Category:Svaman]] on the page before you save it to make it part of the Svaman wiki (preload can be enabled to automate this task, by clicking this link and saving that page. Afterwards, you may need to purge this page, if you still see this message).
blablabkla hekgerlhgerjgheglñeehtfkerlñ


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"When you're telling your own story, it's hard to know where to start." Brandon Stanton has dedicated his life to telling other people's stories on his blog Humans of New York, or HONY, as it's known to its nearly 17 million followers. What began as an idea to photograph 10,000 people on the streets of New York City has led to a wildly successful Facebook page. Not to mention three New York Times best-selling photography books, a 50-day tour of 10 countries sponsored by the United Nations in 2014, visits to the Oval Office with students and educators featured on the blog, and millions of dollars raised for causes brought to light through people's stories.
On Feb. 9, Stanton told his story in Battell Chapel as a Poynter Fellow in Journalism. He started with his windy path to success, explaining, "I was a History major at the University of Georgia; I didn't go to Yale. Then I flunked out of Georgia, went to community college, got my life back on track, and then—just through this connection I had—I was offered a very prestigious job in Chicago trading bonds." Stanton recalled obsessing over his work while he was a bond trader. "I spent two years thinking about nothing but making money," he said. Then, after he was fired, Stanton experienced what he calls "the most momentous moment of my life."
"I made the decision that from here on out I'm going to try to make just enough money where I can do what I want with my time," Stanton said. "That was a real profound reorientation of perspective for me." The key for Stanton was to stop viewing time as a means to obtain other resources and instead to look at time as a resource in itself.
What Stanton enjoyed doing most with his free time was taking pictures. "The whole reason why I'd gotten my first camera was to relieve the stress of work," he said. One day, he took a photo of two young boys and their mothers riding the subway together. Looking back at the candid shot, Stanton realized that if he could continue to approach people and ask to take their pictures, he could create something unique. Not many people approach strangers on the street. While he never harbored hopes of becoming the best photographer in the world, he had always considered himself creative. So he started approaching strangers and asking to take their pictures constantly. He began in Chicago, and then traveled to a few other cities to take portraits before he decided to take his project to New York.
The original concept for HONY was simple: Stanton would move from Chicago to New York City in order to take 10,000 portraits of people on the streets and plot them on a map. When he got to New York in 2010, just as he obsessed over trading, Stanton started obsessing over his photography. When he first moved to the city, he only ate two meals a day and didn't go out to clubs or bars. He spent his time taking photos all day, every day.
Originally, Stanton only took portraits, which he posted with very minimal captions, if any caption at all. Stanton first posted a quote alongside a photo for a picture of a woman wearing only green. He was unsure about posting the image, which he described as nothing special, but recalled that the woman said something interesting: she used to go through stages of wearing different colors, until she discovered that she was happiest in green. "I've been green for 15 years," she told Stanton. He decided to put the photo up with her comment. The photo received significantly more attention than any of the previous posts, and Stanton took note.
Almost six years later, Stanton emphasizes that the blog as it is today is entirely different from the blog that he set out to create. "Humans of New York today is not really about photography," he said. "It's about telling stories." The blog has evolved to become a platform for marginalized voices to be heard. Stanton has traveled to Jordan and Turkey in order to interview Syrian refugees and recently released a series of stories featuring inmates in five federal prisons across the Northeast.
Today, Stanton interviews his subjects for 45 minutes to an hour and then takes their picture afterwards. He's often asked how he gets people to share such personal experiences with him. He explained, "The reason why Humans of New York has so many followers, why it has such an impact, is that through approaching thousands of people, I learned to do it in a way where I can create a moment of intimacy, compassion, whatever you want to call it." He calls this environment the "bubble," in which people feel genuinely heard and validated.
Stanton elaborated on why people may feel compelled to share in the "bubble," saying, "There's something cathartic and easy about telling your story to someone who has no preconceptions of you against which to weigh your opinions or judge your experiences." Stanton noted that he might be the first person that someone has talked to seriously for months. And not only does he talk to his subjects, he really listens to what they have to say.
On the blog, stories are told through captions posted with portraits. Sometimes, a story will be told through multiple captions and different portraits. Many posts go viral—some of the most popular photos have hundreds of thousands of "likes." President Obama even commented on a portrait of a Syrian refugee, a man known as "The Scientist" to HONY followers, welcoming him to the United States.
One audience member asked whether the stories of the thousands of people that Stanton has interviewed ever blur together for him. "You do start to notice themes," conceded Stanton. For example, the 20 year old whose goal is to travel and whose biggest struggle is figuring out what to do with his or her life—"I've had that conversation a thousand times," he said. The key is to discover the telling details that make each person's story unique. "As an interviewer and a storyteller," he explained, "my entire interview process is aimed at getting through that blur."
Stanton's ability to connect with people makes his content unique, but the format on which he shares it is similarly compelling. Why is it that HONY has flourished on Facebook, a social media platform that is essentially made up of people captioning and posting photos of themselves? After all, HONY does something really similar, doesn't it? Turns out it doesn't. The things that people reveal to Stanton aren't the things that people would post on their own profiles—and that's exactly why it's so popular. In an online space that can seem like a never-ending highlight reel, it's refreshing to see the truth of people's stories come to light through Stanton's Facebook posts.
Towards the end of the talk, I couldn't stop thinking about how Stanton was positioning himself for the audience in Battell while telling his story. At times, he lapsed into giving us the highlight-reel version of HONY—and understandably so—There are a lot of highlights to choose from – he's seen so many incredible things, heard so many amazing stories, impacted so many people's lives, and visited so many countries.
But he also made a point to get through the blur of his own successes and reflect on the low points he faced early on. As cheesy as it was, one of the most powerful moments of the talk came when he addressed the students in the audience about failing. He ventured that many in the audience may be so used to success that a fear of failure holds them back. He warns against that, reflecting that if he had feared failure six years ago, "I would have tried to plan all the risk out of it." Without risk, HONY would never have become the personal yet massive platform it is now.


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About Us
Check out the detailed profiles for our available dogs at www.albertaherdingdogrescue.com
Each dog in foster care has their own picture album with a more detailed write up on their history as we know it, the behaviours that we are seeing in foster care and the type of home we think would best suit them.
If you are interested in one of our dogs, please email us for an adoption questionnaire. This must be filled out as completely and truthfully as possible and include references.
We will consider long distance adoptions but we will not ship our dogs to people or places unknown. We require that adopters travel to meet our dogs, once all steps of the process have been completed.
We are a small, nationally registered Not-for-Profit organization based out of Alberta, Canada. Our main source of funding is by donation, from friends and family, or from hosting small agility trials or herding clinics. Regretfully, we are not (yet) a registered charity so we cannot issue tax receipts.
Our volunteer network is small and knowledgeable foster homes are hard to come by. The status of these resources at any given time dictate the number of dogs we can help out.
We would appreciate your support, either by direct donation through PayPal or through our GoFundMe Wish List campaign (https://www.gofundme.com/AB-HerdingDogRescue).
We would rather spend time helping our dogs become better pets than bombard you with soliciting emails or facebook messages. So any donation amount, any time is greatly appreciated.
AB-HDR is located approximately 1.5 hours NW of Calgary, Alberta, Canada.
We are not a shelter with regular office or store hours. Although many of our dogs are located at Cavorting K9s Retreat, all our dogs are fostered in private homes. All visits are by appointment with the foster family.


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Butcher and war veteran James Dunning passes away
James Dunning, the writer, distinguished war soldier and former butcher, has died at the age of 95.
A contributor to Meat Trades Journal for more than 20 years, his books included the 1988 publication 'A Century of Service', which chronicled the first 100 years of the Journal or, as he put it, 'The story of the Meat Trades Journal and Cattle Salesmens Gazette' (the organ of the British, Colonial and Foreign Live Stock and Dead Meat Trades) 1888-1988'.
Former MTJ editor Fred A'Court said "As with many brave wartime soldiers Jimmy Dunning came across as a modest, quiet and courteous man, who never mentioned his numerous roles as a commando during the Second World War, including one of the few successful attacks in the disastrous 1942 Dieppe invasion, which saw 60% of the commandoes killed, wounded or taken prisoner, and the loss of 108 RAF aircraft, a Royal Navy destroyer and 33 assault craft. His obituary in The Times gives a detailed account of his wartime service and the books he wrote about commando training and his wartime experiences."
Dunning came from a family of butchers and he returned to the trade after leaving the army in 1958. In 1962 he was president of the Southampton Association of Meat Traders. Among his specially written features were centenary supplements on Smithfield Market and the New Zealand meat trade with Britain. Other articles included features on butchers in the army, on the liner QE11 and on Harrods Food Hall. He also wrote about butchers in Cyprus, Malta, France, Italy, Greece and even the Falkland Islands.
For several years he contributed regularly to the 'Country Butcher's Diary'. In 1985 he wrote what was probably his first full-length book 'Britain's Butchers - The Trade Through The Ages' for Meat Trades Journal publications.
On completing 'A Century of Service' he admitted he would have liked twice as much time to research and write it, but the centenary couldn't be put off!
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