Silva Erica, Michaca Manuel, Munson Brenton, Bean Gordon J, Jaeger Philipp A, Licon Katherine, Winzeler Elizabeth A, Ideker Trey
Department of Medicine, University of California, San Diego, La Jolla, California
Department of Medicine, University of California, San Diego, La Jolla, California.
G3 (Bethesda). 2020 Sep 2;10(9):2981-2988. doi: 10.1534/g3.120.401417.
Genetic screens in have allowed for the identification of many genes as sensors or effectors of DNA damage, typically by comparing the fitness of genetic mutants in the presence or absence of DNA-damaging treatments. However, these static screens overlook the dynamic nature of DNA damage response pathways, missing time-dependent or transient effects. Here, we examine gene dependencies in the dynamic response to ultraviolet radiation-induced DNA damage by integrating ultra-high-density arrays of 6144 diploid gene deletion mutants with high-frequency time-lapse imaging. We identify 494 ultraviolet radiation response genes which, in addition to recovering molecular pathways and protein complexes previously annotated to DNA damage repair, include components of the CCR4-NOT complex, tRNA wobble modification, autophagy, and, most unexpectedly, 153 nuclear-encoded mitochondrial genes. Notably, mitochondria-deficient strains present time-dependent to ultraviolet radiation, posing impaired mitochondrial function as a protective factor in the ultraviolet radiation response.
酵母中的遗传筛选已使许多基因被鉴定为DNA损伤的传感器或效应器，通常是通过比较存在或不存在DNA损伤处理时遗传突变体的适应性来实现的。然而，这些静态筛选忽略了DNA损伤反应途径的动态性质，遗漏了时间依赖性或瞬时效应。在这里，我们通过将6144个二倍体基因缺失突变体的超高密度阵列与高频延时成像相结合，研究了对紫外线诱导的DNA损伤的动态反应中的基因依赖性。我们鉴定出494个紫外线反应基因，除了恢复先前注释为DNA损伤修复的分子途径和蛋白质复合物外，还包括CCR4-NOT复合物的成分、tRNA摆动修饰、自噬，最出乎意料的是，还有153个核编码的线粒体基因。值得注意的是，线粒体缺陷菌株对紫外线呈现出时间依赖性的敏感性，表明线粒体功能受损是紫外线反应中的一种保护因素。
