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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Proteome-wide identification of in vivo targets of DNA damage checkpoint kinases
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Proteome-wide identification of in vivo targets of DNA damage checkpoint kinases

机译：蛋白质组学鉴定DNA损伤检查点激酶的体内靶标

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摘要

Understanding the role of DNA damage checkpoint kinases in the cellular response to genotoxic stress requires the knowledge of their substrates. Here, we report the use of quantitative phospho-proteomics to identify in vivo kinase substrates of the yeast DNA damage checkpoint kinases Mec1, Tel1, and Rad53 (orthologs of human ATR, ATM, and CHK2, respectively). By analyzing 2,689 phosphorylation sites in wild-type and various kinase-null cells, 62 phosphorylation sites from 55 proteins were found to be controlled by the DNA damage checkpoint. Examination of the dependency of each phosphorylation on Mec1 and Tel1 or Rad53, combined with sequence and biochemical analysis, revealed that many of the identified targets are likely direct substrates of these kinases. In addition to several known targets, 50 previously unde-scribed targets of the DNA damage checkpoint were identified, suggesting that a wide range of cellular processes is likely regulated by Mec1, Tel1, and Rad53.
机译：要了解DNA损伤检查点激酶在细胞对遗传毒性胁迫的反应中的作用，需要了解其底物。在这里，我们报告使用定量磷酸化蛋白质组学来确定酵母DNA损伤检查点激酶Mec1，Tel1和Rad53（分别为人ATR，ATM和CHK2的直系同源物）的体内激酶底物。通过分析野生型和各种激酶无效细胞中的2689个磷酸化位点，发现55种蛋白质中的62个磷酸化位点受DNA损伤检查点控制。结合序列和生化分析，对每个磷酸化对Mec1和Tel1或Rad53的依赖性进行的检查显示，许多已鉴定的靶标可能是这些激酶的直接底物。除了几个已知的靶标外，还鉴定了50个DNA损伤检查点以前未描述的靶标，这表明Mec1，Tel1和Rad53可能调节了广泛的细胞过程。

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