Double-strand breaks that are induced post-replication trigger establishment of damage-induced cohesion in Saccharomyces cerevisiae, locally at the break-site and genome wide on undamaged chromosomes. The translesion synthesis polymerase, polymerase η, is required for generation of damage-induced cohesion genome wide. However, its precise role and regulation in this process is unclear. Here, we investigated the possibility that the cyclin dependent kinase Cdc28 and the acetyltransferase Eco1 modulate polymerase η activity. Through in vitro phosphorylation and structure modeling, we showed that polymerase η is an attractive substrate for Cdc28. Mutation of the putative Cdc28-phosphorylation site Ser14 to Ala, not only affected polymerase η protein level, but also prevented generation of damage-induced cohesion in vivo We also demonstrated that Eco1 acetylated polymerase η in vitro Certain non-acetylatable polymerase η mutants showed reduced protein level, deficient nuclear accumulation and increased ultraviolet irradiation sensitivity. In addition, we found that both Eco1 and subunits of the cohesin network are required for cell survival after ultraviolet irradiation. Our findings support functionally important Cdc28-mediated phosphorylation, as well as posttranslational modifications of multiple lysine residues that modulate polymerase η activity, and provide new insights into understanding the regulation of polymerase η for damage-induced cohesion.

中文翻译：

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DNA 聚合酶 η 的翻译后调节，与酿酒酵母损伤诱导的内聚力的联系。


复制后诱导的双链断裂触发了酿酒酵母中损伤诱导的内聚力的建立，局部在断裂位点和未受损染色体上的基因组范围内。跨损伤合成聚合酶聚合酶 η 是在基因组范围内产生损伤诱导的内聚力所必需的。然而，其在此过程中的确切作用和调节尚不清楚。在这里，我们研究了细胞周期蛋白依赖性激酶 Cdc28 和乙酰转移酶 Eco1 调节聚合酶 η 活性的可能性。通过体外磷酸化和结构建模，我们表明聚合酶 η 是 Cdc28 的一个有吸引力的底物。假定的 Cdc28 磷酸化位点 Ser14 突变为 Ala，不仅影响聚合酶 η 蛋白水平，而且还阻止了体内损伤诱导的内聚力的产生。蛋白质水平、核积累不足和紫外线照射敏感性增加。此外，我们发现Eco1和粘连蛋白网络的亚基都是紫外线照射后细胞存活所必需的。我们的研究结果支持功能上重要的 Cdc28 介导的磷酸化，以及调节聚合酶 η 活性的多个赖氨酸残基的翻译后修饰，并为理解聚合酶 η 对损伤诱导的内聚力的调节提供了新的见解。