Lsm12 Mediates Deubiquitination of DNA Polymerase η To Help Saccharomyces cerevisiae Resist Oxidative Stress
Lsm12 Mediates Deubiquitination of DNA Polymerase η To Help Saccharomyces cerevisiae Resist Oxidative Stress
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Lsm12 介导 DNA 聚合酶 eta 去泛素化，帮助酿酒酵母抵抗氧化应激
DOI：
10.1128/aem.01988-18
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发表时间：
2019-01-01
影响因子：
4.4
通讯作者：
Wu, Jing
中科院分区：
文献类型：
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作者：
Yao, Rui;Shi, Liujia;Wu, Jing
在酿酒酵母中，Y 家族 DNA 聚合酶 eta (Pol eta) 通过跨损伤 DNA 合成来调节基因组稳定性，以响应不同形式的环境应激。为了阐明Poleta在氧化应激诱导的DNA损伤中的作用，我们删除或过表达相应的基因RAD30，并利用转录组分析筛选与RAD30相关的潜在基因，以响应DNA损伤。在2 mM H2O2处理下，与野生型菌株相比，RAD30的缺失导致存活率下降2.2倍，DNA损伤增加2.8倍，而RAD30的过表达使存活率增加1.2倍，DNA损伤减少1.4倍。转录组和表型分析确定 Lsm12 是参与氧化应激诱导的 DNA 损伤的主要因素。删除 LSM12 会导致生长缺陷，而其过表达在 2 mM H2O2 处理下会增强细胞生长。这种效应是由于 Lsm12 与 Pol eta 的 UBZ 结构域发生物理相互作用，通过 Ubp3 增强 Pol eta 去泛素化，从而促进 Pol eta 招募。总的来说，这些发现表明 Lsm12 是一种介导 Pol eta 去泛素化的新型调节剂，以促进其在氧化应激下的募集。此外，这项研究提供了一种在发酵过程中维持工业菌株基因组稳定性的潜在策略。 重要性 Pol eta被证明对酿酒酵母的细胞生长至关重要，其相应基因RAD30的缺失在暴露于2 mM H2O2的氧化应激下会导致严重的生长缺陷。此外，我们发现 Lsm12 与 Pol eta 发生物理相互作用，并促进 Pol eta 去泛素化和招募。总体而言，这些发现表明 Lsm12 是一种介导 Pol eta 去泛素化的新型调节因子，可调节其募集以响应氧化应激引起的 DNA 损伤。
In Saccharomyces cerevisiae, the Y family DNA polymerase eta (Pol eta) regulates genome stability in response to different forms of environmental stress by translesion DNA synthesis. To elucidate the role of Pol eta in oxidative stress-induced DNA damage, we deleted or overexpressed the corresponding gene RAD30 and used transcriptome analysis to screen the potential genes associated with RAD30 to respond to DNA damage. Under 2 mM H2O2 treatment, the deletion of RAD30 resulted in a 2.2-fold decrease in survival and a 2.8-fold increase in DNA damage, whereas overexpression of RAD30 increased survival and decreased DNA damage by 1.2- and 1.4-fold, respectively, compared with the wild-type strain. Transcriptome and phenotypic analyses identified Lsm12 as a main factor involved in oxidative stress-induced DNA damage. Deleting LSM12 caused growth defects, while its over-expression enhanced cell growth under 2 mM H2O2 treatment. This effect was due to the physical interaction of Lsm12 with the UBZ domain of Pol eta to enhance Pol eta deubiquitination through Ubp3 and consequently promote Pol eta recruitment. Overall, these findings demonstrate that Lsm12 is a novel regulator mediating Pol eta deubiquitination to promote its recruitment under oxidative stress. Furthermore, this study provides a potential strategy to maintain the genome stability of industrial strains during fermentation.IMPORTANCE Pol eta was shown to be critical for cell growth in the yeast Saccharomyces cerevisiae, and deletion of its corresponding gene RAD30 caused a severe growth defect under exposure to oxidative stress with 2 mM H2O2. Furthermore, we found that Lsm12 physically interacts with Pol eta and promotes Pol eta deubiquitination and recruitment. Overall, these findings indicate Lsm12 is a novel regulator mediating Pol eta deubiquitination that regulates its recruitment in response to DNA damage induced by oxidative stress.