Identification of Cables1 as a critical host factor that promotes ALV-J replication via genome-wide CRISPR/Cas9 gene knockout screening

文献类型： 外文期刊

第一作者： Liu, Peng

作者： Liu, Peng;Chen, Yuntong;Wang, Suyan;Yu, Mengmeng;Liu, Yongzhen;Guo, Ru;Zhang, Li;Xu, Zhuangzhuang;Wang, Caiying;Qi, Xiaole;Zhang, Yanping;Cui, Hongyu;Duan, Yulu;Gao, Yulong;Jiang, Jinghua;Gao, Fei;Wang, Caiying;Wu, Sen;Wu, Sen;Gao, Yulong;Gao, Yulong

作者机构：

期刊名称：JOURNAL OF BIOLOGICAL CHEMISTRY （ 影响因子：3.9； 五年影响因子：4.3 ）

ISSN：

年卷期： 2024 年 300 卷 11 期

页码：

收录情况： SCI

摘要： Avian leukosis virus subgroup J (ALV-J), a member of the genus Alpharetrovirus, possesses a small genome and exploits a vast array of host factors during its replication cycle. To identify host factors required for ALV-J replication and potentially guide the development of key therapeutic targets for ALV-J prevention, we employed a chicken genome-wide CRISPR/Cas9 knockout library to screen host factors involved in ALV-J infection within DF-1 cells. This screening revealed 42 host factors critical for ALV-J infection. Subsequent knockout assays showed that the absence of the genes encoding cycle-regulatory proteins, namely, Cables1, CDK1, and DHFR, significantly inhibited ALV-J replication. Notably, Cables1 knockout cell lines displayed the most pronounced inhibitory effect. Conversely, overexpression assays confirmed that Cables1 significantly promotes ALV-J replication. Immunoprecipitation assays further indicated that Cables1 specifically interacts with the viral protein p15 (viral protease) among all ALV-J proteins, enhancing ALV-J p15 polyubiquitination. Additionally, we identified 26 lysine residues of ALV-J p15 as key sites for ubiquitination, and their replacement with arginine attenuated the replication ability of ALV-J in both in vitro and in vivo assays. This study demonstrates that Cables1 is a critical replication-dependent host factor of ALV-J by enhancing p15 ubiquitination and thereby promoting viral replication. Overall, these fi ndings contribute to a deeper understanding of the ALJ-V replication mechanism and offer a potential target for the prevention and control of ALV-J infection.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序