Sánchez Arancha, Russell Paul
Department of Cell and Molecular Biology, The Scripps Research Institute, La Jolla, California, United States of America.
PLoS One. 2015 May 12;10(5):e0126598. doi: 10.1371/journal.pone.0126598. eCollection 2015.
DNA replication errors are a major source of genome instability in all organisms. In the fission yeast Schizosaccharomyces pombe, the DNA damage response protein Brc1 binds phospho-histone H2A (γH2A)-marked chromatin during S-phase, but how Brc1 protects genome integrity remains unclear. Here we report that the non-homologous end-joining (NHEJ) protein Ku becomes critical for survival of replication stress in brc1∆ cells. Ku's protective activity in brc1∆ cells does not involve its canonical NHEJ function or its roles in protecting telomeres or shielding DNA ends from Exo1 exonuclease. In brc1∆ pku80∆ cells, nuclear foci of Rad52 homologous recombination (HR) protein increase and Mus81-Eme1 Holliday junction resolvase becomes critical, indicating increased replication fork instability. Ku's localization at a ribosomal DNA replication fork barrier associated with frequent replisome-transcriptosome collisions increases in brc1∆ cells and increased collisions correlate with an enhanced requirement for Brc1. These data indicate that Ku stabilizes replication forks in the absence of Brc1.
DNA复制错误是所有生物体基因组不稳定的主要来源。在裂殖酵母粟酒裂殖酵母中,DNA损伤反应蛋白Brc1在S期结合磷酸化组蛋白H2A(γH2A)标记的染色质,但Brc1如何保护基因组完整性仍不清楚。在此,我们报道非同源末端连接(NHEJ)蛋白Ku对brc1∆细胞中复制应激的存活至关重要。Ku在brc1∆细胞中的保护活性不涉及其典型的NHEJ功能,也不涉及其在保护端粒或保护DNA末端免受Exo1核酸外切酶作用方面的作用。在brc1∆ pku80∆细胞中,Rad52同源重组(HR)蛋白的核灶增加,Mus81-Eme1 Holliday连接解离酶变得至关重要,表明复制叉不稳定性增加。在brc1∆细胞中,Ku在与频繁的复制体-转录体碰撞相关的核糖体DNA复制叉屏障处的定位增加,并且增加的碰撞与对Brc1的需求增加相关。这些数据表明,在没有Brc1的情况下,Ku稳定复制叉。