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. 2009 Aug 15;8(16):2664-6.
doi: 10.4161/cc.8.16.9367. Epub 2009 Aug 26.

Cell cycle-dependent deacetylation of telomeric histone H3 lysine K56 by human SIRT6

Eriko Michishita, Ronald A McCord, Lisa D Boxer, Matthew F Barber, Tao Hong, Or Gozani, Katrin F Chua

Cell cycle-dependent deacetylation of telomeric histone H3 lysine K56 by human SIRT6

Eriko Michishita et al. Cell Cycle. .
No abstract available

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Figures

Figure 1
Figure 1
Evidence for global and telomere-specific regulalion of H3K56Ac by human SIRT6. (A) Mass spedrometry analysis showing in vilro deacelylation by SIRT6 of an acelylaled H3K56 peplide (GTVALREIRRYQK(Ac)STELLIRK), shown by the appearance of a SIRT6-dependent peak (arrow) 42 Da smaller than the acetylaled peptide Molecular weighls (Da) of the acetylated and deacetylated peptide peaks are indicated. NAD-dependent deacelylation reactions were performed as previously described. (B) Mass spectrometry as in (A) showing in vitro deacetylation of H3K56Ac peptide by SIRT 1. (C) Western analysis of whole cell extracts from cells expressing Flag-tagged wild-type SIRT6 (SIRT6-WT), calalytically inactive SIRT6 (SIRT6-HY), or empty vector control (pcDNA). (D) Western analysis of whole cell extracts from cells expressing Flag-tagged SIRT6 or SIRT1 compared to pcDNA control. (E) Western analysis of global H3K56Ac levels in SIRT6 knockdown (S6KD1 and S6KD2) or control (pSR) cells following cell cycle arrest due to thymidine, colcemid or nocodazole blocks. The corresponding cell cycle phases enriched in these cultures are indicated. (F) Telomere ChIP analysis showing hyperacetylation of H3K56 at telomeres in S-phase-enriched U2OS cell cultures. IgG ChIP and input DNA are present as controls. (G) Quantification of Telomere ChIP analyses in U2OS cells at 4 and 12 hours following release from a thymidine block, corresponding to S and G2/M phases, respectively. In (C-G), antibodies were αH3K56Ac (Epitomics); αflag (Sigma), αH3 (Abcam); αSIRT6 antibodies were previously described.
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