Department of Biochemistry and Biophysics, School of Basic Medical Sciences, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Beijing Key Laboratory of Protein Posttranslational Modifications and Cell Function, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China.
Jingjie PTM BioLab (Hangzhou) Co. Ltd., Hangzhou 310018, China.
Sci Adv. 2020 Mar 13;6(11):eaay4697. doi: 10.1126/sciadv.aay4697. eCollection 2020 Mar.
Previously, we reported that chromodomain Y-like (CDYL) acts as a crotonyl-coenzyme A hydratase and negatively regulates histone crotonylation (Kcr). However, the global CDYL-regulated crotonylome remains unclear. Here, we report a large-scale proteomics analysis for protein Kcr. We identify 14,311 Kcr sites across 3734 proteins in HeLa cells, providing by far the largest crotonylome dataset. We show that depletion of CDYL alters crotonylome landscape affecting diverse cellular pathways. Specifically, CDYL negatively regulated Kcr of RPA1, and mutation of the Kcr sites of RPA1 impaired its interaction with single-stranded DNA and/or with components of resection machinery, supporting a key role of RPA1 Kcr in homologous recombination DNA repair. Together, our study indicates that protein crotonylation has important implication in various pathophysiological processes.
此前，我们报道了 chromodomain Y-like（CDYL）作为丙二酰辅酶 A 水合酶发挥作用，负调控组蛋白丙二酰化（Kcr）。然而，CDYL 调控的丙二酰化组蛋白组仍不清楚。在这里，我们报告了一个大规模的蛋白质组学分析用于蛋白质 Kcr。我们在 HeLa 细胞中鉴定了 3734 种蛋白质中的 14311 个 Kcr 位点，提供了迄今为止最大的丙二酰化组蛋白数据集。我们表明，CDYL 的耗竭改变了丙二酰化组蛋白图谱，影响了多种细胞途径。具体来说，CDYL 负调控 RPA1 的 Kcr，并且 RPA1 的 Kcr 位点突变会损害其与单链 DNA 的相互作用和/或与切除机制的成分的相互作用，支持 RPA1 Kcr 在同源重组 DNA 修复中的关键作用。总之，我们的研究表明蛋白质丙二酰化在各种病理生理过程中具有重要意义。
