ATM and SIRT6/SNF2H Mediate Transient H2AX Stabilization When DSBs Form by Blocking HUWE1 to Allow Efficient γH2AX Foci Formation

• Published in: Cell reports (Cambridge) 2015-12, Vol.13 (12), p.2728-2740
• Main Authors: Atsumi, Yuko, Minakawa, Yusuke, Ono, Masaya, Dobashi, Sachiko, Shinohe, Keitaro, Shinohara, Akira, Takeda, Shunichi, Takagi, Masatoshi, Takamatsu, Nobuhiko, Nakagama, Hitoshi, Teraoka, Hirobumi, Yoshioka, Ken-ichi
• Format: Article
• Language: English
• Subjects: Animals; Ataxia Telangiectasia Mutated Proteins - metabolism; Chromatin - metabolism; DNA Breaks, Double-Stranded; HeLa Cells; Histones - genetics; Histones - metabolism; Humans; Mice; Phosphorylation; Sirtuins - genetics; Sirtuins - metabolism; Tumor Suppressor Proteins; Ubiquitin-Protein Ligases - antagonists & inhibitors; Ubiquitin-Protein Ligases - metabolism
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2015.11.054

In response to DNA double-strand breaks (DSBs), H2AX is rapidly phosphorylated at Ser139 to promote DSB repair. Here we show that H2AX is rapidly stabilized in response to DSBs to efficiently generate γH2AX foci. This mechanism operated even in quiescent cells that barely expressed H2AX. H2AX stabilization resulted from the inhibition of proteasome-mediated degradation. Synthesized H2AX ordinarily underwent degradation through poly-ubiquitination mediated by the E3 ligase HUWE1; however, H2AX ubiquitination was transiently halted upon DSB formation. Such rapid H2AX stabilization by DSBs was associated with chromatin incorporation of H2AX and halting of its poly-ubiquitination mediated by the ATM kinase, the sirtuin protein SIRT6, and the chromatin remodeler SNF2H. H2AX Ser139, the ATM phosphorylation site, was essential for H2AX stabilization upon DSB formation. Our results reveal a pathway controlled by ATM, SIRT6, and SNF2H to block HUWE1, which stabilizes H2AX and induces its incorporation into chromatin only when cells are damaged. [Display omitted] •H2AX is degraded via poly-ubiquitination mediated by HUWE1•H2AX is transiently stabilized upon DSB formation to efficiently form γH2AX foci•H2AX stabilization is mediated by ATM, SIRT6, and SNF2H Atsumi et al. show that H2AX is transiently stabilized upon DSB formation. H2AX stabilization is mediated by the ATM kinase, sirtuin protein SIRT6, and ISWI family chromatin remodeler SNF2H. This enables efficient γH2AX foci formation and DSB repair even in the quiescent cellular state in which cells rarely express H2AX.