the swr1 histone replacement complex causes genetic instability and genome-wide transcription misregulation in the absence of h2a.zswr1组蛋白替换复杂引起遗传不稳定和全基因组转录misregulation h2a.z的缺失.pdfVIP

The SWR1 Histone Replacement Complex Causes Genetic Instability and Genome-Wide Transcription Misregulation in the Absence of H2A.Z 1. 1. ´ ´ 2 ´ 1 Macarena Morillo-Huesca , Marta Clemente-Ruiz , Eloısa Andujar , Felix Prado * 1 Department of Molecular Biology, CABIMER-CSIC, Seville, Spain, 2 Genomics Unit, CABIMER-CSIC, Seville, Spain Abstract The SWR1 complex replaces the canonical histone H2A with the variant H2A.Z (Htz1 in yeast) at specific chromatin regions. This dynamic alteration in nucleosome structure provides a molecular mechanism to regulate transcription, gene silencing, chromosome segregation and DNA repair. Here we show that genetic instability, sensitivity to drugs impairing different cellular processes and genome-wide transcriptional misregulation in htz1D can be partially or totally suppressed if SWR1 is not formed (swr1D), if it forms but cannot bind to chromatin (swc2D) or if it binds to chromatin but lacks histone replacement activity (swc5D and the ATPase-dead swr1-K727G). These results suggest that in htz1D the nucleosome remodelling activity of SWR1 affects chromatin integrity because of an attempt to replace H2A with Htz1 in the absence of the latter. This would impair transcription and, either directly or indirectly, other cellular processes. Specifically, we show that in htz1D, the SWR1 complex causes an accumulation of recombinogenic DNA damage by a mechanism dependent on phosphorylation of H2A at Ser129, a modification that occurs in response to DNA damage, suggesting that the SWR1 complex impairs the repair of spontaneous DNA damage in htz1D. In addition, SWR1 causes DSBs sensitivity in htz1D; consistently, in t