Department of Biochemistry and Biophysics, Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, CA 94158, USA; Tetrad Graduate Program, University of California, San Francisco, San Francisco, CA 94158, USA.
Department of Biochemistry and Biophysics, Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, CA 94158, USA.
Mol Cell. 2021 Mar 18;81(6):1319-1336.e9. doi: 10.1016/j.molcel.2021.01.014. Epub 2021 Feb 3.
The human ubiquitin proteasome system, composed of over 700 ubiquitin ligases (E3s) and deubiquitinases (DUBs), has been difficult to characterize systematically and phenotypically. We performed chemical-genetic CRISPR-Cas9 screens to identify E3s/DUBs whose loss renders cells sensitive or resistant to 41 compounds targeting a broad range of biological processes, including cell cycle progression, genome stability, metabolism, and vesicular transport. Genes and compounds clustered functionally, with inhibitors of related pathways interacting similarly with E3s/DUBs. Some genes, such as FBXW7, showed interactions with many of the compounds. Others, such as RNF25 and FBXO42, showed interactions primarily with a single compound (methyl methanesulfonate for RNF25) or a set of related compounds (the mitotic cluster for FBXO42). Mutation of several E3s with sensitivity to mitotic inhibitors led to increased aberrant mitoses, suggesting a role for these genes in cell cycle regulation. Our comprehensive CRISPR-Cas9 screen uncovered 466 gene-compound interactions covering 25% of the interrogated E3s/DUBs.
人类泛素蛋白酶体系统由超过 700 种泛素连接酶（E3s）和去泛素化酶（DUBs）组成，其系统和表型特征一直难以进行全面的描述。我们进行了化学遗传学 CRISPR-Cas9 筛选，以鉴定那些缺失会导致细胞对靶向广泛生物过程的 41 种化合物（包括细胞周期进程、基因组稳定性、代谢和囊泡运输）敏感或耐药的 E3s/DUBs。基因和化合物在功能上聚类，相关途径的抑制剂与 E3s/DUBs 以相似的方式相互作用。一些基因，如 FBXW7，与许多化合物表现出相互作用。其他基因，如 RNF25 和 FBXO42，则主要与单个化合物（RNF25 的甲基甲磺酸酯）或一组相关化合物（FBXO42 的有丝分裂簇）表现出相互作用。对几种对有丝分裂抑制剂敏感的 E3s 的突变导致异常有丝分裂增加，表明这些基因在细胞周期调控中发挥作用。我们全面的 CRISPR-Cas9 筛选揭示了 466 个基因-化合物相互作用，涵盖了被检测的 E3s/DUBs 的 25%。
