A genome-wide CRISPR screen identifies HuR as a regulator of apoptosis induced by dsRNA and virus.
A genome-wide CRISPR screen identifies HuR as a regulator of apoptosis induced by dsRNA and virus.
复制标题
全基因组 CRISPR 筛选将 HuR 鉴定为 dsRNA 和病毒诱导的细胞凋亡的调节因子。
DOI：
10.1242/jcs.258855
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发表时间：
2022
影响因子：
4
通讯作者：
Ping Zhang
中科院分区：
文献类型：
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作者：
Huixin Gao;Yuxia Lin;Changbai Huang;Xiaobo Li;M. Diamond;Chao Liu;Rong Zhang;Ping Zhang
我们在A549细胞中进行了无偏倚的全基因组CRISPR/Cas9筛选，以确定参与dsRNA触发的细胞死亡的潜在调控因子。在几个候选基因中，我们发现了编码HU抗原R(HUR)的RNA结合蛋白ELAV样蛋白1(ELAVL1)。HUR的耗尽导致dsRNA诱导的细胞死亡减少。HUR主要参与细胞凋亡，其所有的RNA识别基序都是其促凋亡功能所必需的。我们进一步表明，HUR缺失对抗凋亡基因BCL2的mRNA水平没有影响，相反，它以一种不依赖帽子的方式下调其翻译。多聚体分离研究表明，HUR延缓了非翻译多聚体池中的BCL2 mRNA的表达。此外，HUR缺失对dsRNA诱导的细胞凋亡的保护作用需要BCL2的存在，这表明HUR的促凋亡功能是通过抑制BCL2来执行的。一直以来，HUR调节脑心肌炎或塞姆利基森林病毒感染诱导的细胞凋亡。总体而言，我们的工作确定了一套调节dsRNA诱导的细胞死亡的蛋白质，并阐明了HUR作为促凋亡因子的机制。
We performed an unbiased whole-genome CRISPR/Cas9 screen in A549 cells to identify potential regulators involved in cell death triggered by dsRNA. Of several top candidate genes, we identified the RNA binding protein ELAV like protein 1 (ELAVL1) that encodes Hu antigen R (HuR). Depletion of HuR led to less cell death induced by dsRNA. HuR is mainly involved in the apoptosis, and all of its RNA recognition motifs are essential for its proapoptotic function. We further showed that the HuR depletion had no influence on the mRNA level of an anti-apoptotic gene, BCL2, instead downregulated its translation in a cap-independent way. Polysome fractionation studies showed that HuR retarded the BCL2 mRNA in the non-translating pool of polysomes. Moreover, protection from dsRNA-induced apoptosis by HuR depletion required the presence of BCL2, indicating that the proapoptotic function of HuR is executed by suppressing BCL2. Consistently, HuR regulated apoptosis induced by infection of encephalomyocarditis or Semliki Forest virus. Collectively, our work identified a suite of proteins that regulate dsRNA-induced cell death, and elucidated the mechanism by which HuR acts as a pro-apoptotic factor.