哺乳动物细胞中表达的细菌Cas2Em蛋白对生长的抑制作用。
Growth inhibition by bacterial Cas2Em proteins expressed in mammalian cells.
作者信息
Yu Xin, Wang Lei, Zou Lin, Li Mengjie, Li Tiansheng, Hou Linlin, Guo Yameng, Shen Danfeng, Sun Guiqin, Qu Di, Cheng Xunjia, Chen Li
机构信息
Department of Medical Microbiology, Key Laboratory of Medical Molecular Virology of Ministries of Education and Health, School of Basic Medical Sciences, Fudan University Shanghai 200032, China.
Roche Innovation Center Shanghai Shanghai 201203, China.
出版信息
Am J Transl Res. 2020 Jun 15;12(6):2499-2520. eCollection 2020.
BACKGROUND
Clustered regularly interspaced short palindromic repeats (CRISPRs) and the CRISPR-associated (Cas) proteins are bacterial adaptive immune system for survival. In our previous study, we demonstrate that polyploid giant bacterial cells (PGBC) induced by Cas2 protein is a step required by new spacer acquisition reaction catalyzed by Cas1/Cas2 complex. We also demonstrated that a carboxyl terminal domain on Cas2Em (the protein Cas2 cloned from ) is sufficient and enough for PGBC. Thus, the potential role of Cas2Em in microbial-host interaction was explored in this study.
METHODS
The impacts of Cas2Em on growth of both CHO-K1 and Hela cells were investigated. The subcellular localization and potential molecular target of Ca2Em were studied.
RESULTS
The growth of mammalian cells were inhibited by Cas2Em protein via G1 arresting and apoptosis. In addition, we also demonstrated that Cas2Em was tightly associated with nuclear outer membrane and could be immunoprecipitated with 14-3-3γ through a 30 amino acid domain (homology of CLK2).
CONCLUSION
Cas2Em significantly suppressed the growth of mammalian cells indicating Cas2 proteins play an important role in mammalian cells.
背景
成簇规律间隔短回文重复序列（CRISPRs）及CRISPR相关（Cas）蛋白是细菌用于生存的适应性免疫系统。在我们之前的研究中，我们证明由Cas2蛋白诱导产生的多倍体巨型细菌细胞（PGBC）是Cas1/Cas2复合体催化的新间隔序列获取反应所需的一个步骤。我们还证明Cas2Em（从 克隆的Cas2蛋白）上的羧基末端结构域对PGBC来说是足够的。因此，本研究探讨了Cas2Em在微生物-宿主相互作用中的潜在作用。
方法
研究了Cas2Em对CHO-K1细胞和Hela细胞生长的影响。研究了Ca2Em的亚细胞定位和潜在分子靶点。
结果
Cas2Em蛋白通过G1期阻滞和凋亡抑制哺乳动物细胞的生长。此外，我们还证明Cas2Em与核外膜紧密相关，并且可以通过一个30个氨基酸的结构域（与CLK2同源）与14-3-3γ进行免疫沉淀。
结论
Cas2Em显著抑制哺乳动物细胞的生长，表明Cas2蛋白在哺乳动物细胞中起重要作用。
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