NSUN2 mediates distinct pathways to regulate enterovirus 71 replication

Lishi Liu; Zhen Chen; Kui Zhang; Haojie Hao; Li Ma; Haizhou Liu; Baocheng Yu; Shuang Ding; Xueyan Zhang; Miao Zhu; Xiang Guo; Yi Liu; Haibin Liu; Fang Huang; Ke Peng; Wuxiang Guan

doi:10.1016/j.virs.2024.05.002

August 2024

Lishi Liu, Zhen Chen, Kui Zhang, Haojie Hao, Li Ma, Haizhou Liu, Baocheng Yu, Shuang Ding, Xueyan Zhang, Miao Zhu, Xiang Guo, Yi Liu, Haibin Liu, Fang Huang, Ke Peng and Wuxiang Guan. NSUN2 mediates distinct pathways to regulate enterovirus 71 replication[J]. Virologica Sinica, 2024, 39(4): 574-586. doi: 10.1016/j.virs.2024.05.002

Citation: Lishi Liu, Zhen Chen, Kui Zhang, Haojie Hao, Li Ma, Haizhou Liu, Baocheng Yu, Shuang Ding, Xueyan Zhang, Miao Zhu, Xiang Guo, Yi Liu, Haibin Liu, Fang Huang, Ke Peng, Wuxiang Guan. NSUN2 mediates distinct pathways to regulate enterovirus 71 replication .VIROLOGICA SINICA, 2024, 39(4) : 574-586. http://dx.doi.org/10.1016/j.virs.2024.05.002

m5C甲基转移酶NSUN2调控EV71复制的机制研究

刘丽诗 ^a,b ,
陈珍 ^a ,
张奎 ^a,b ,
郝好杰 ^a ,
马力 ^a,b ,
刘海舟 ^a ,
余宝成 ^a,b ,
丁爽 ^a ,
张雪艳 ^a ,
朱妙 ^a,b ,
郭祥 ^a,b ,
刘艺 ^c ,
刘海滨 ^a ,
黄方 ^c,, ,
彭珂 ^a,c,, ,
关武祥 ^a,c,,

cstr: 32224.14.j.virs.2024.05.002

a. 中国科学院生物安全大科学研究中心，武汉病毒所新发传染病研究中心;
b. 中国科学院大学;
c. 湖北江夏实验室

通讯作者： 黄方, huangf@wh.iov.cn; 彭珂, pengke@wh.iov.cn; 关武祥, guanwx@wh.iov.cn
收稿日期： 2024-03-03
录用日期： 2024-04-03

摘要

RNA的转录后修饰在真核生物中非常常见，到目前为止报道的RNA修饰有170多种。其中，m5C修饰已被证实存在于tRNA，rRNA和mRNA中，用于调控RNA结构和功能。越来越多的证据表明，甲基转移酶NSUN2催化病毒 RNA 上的m5C修饰对不同病毒的复制至关重要。尽管m5C修饰对于RNA代谢的一些功能已被证实，但NSUN2在调控病毒复制方面的其他潜在作用在很大程度上仍不为人知。在本研究中，我们发现EV71基因组上存在m5C修饰，并在EV71感染后m5C甲基转移酶NSUN2表达显著上调，其定位从细胞核转移至细胞质。我们证实了NSUN2是催化EV71 RNA上m5C修饰的甲基转移酶，能促进EV71的复制以及病毒RNA核糖体的加载，敲低NUSN2抑制了EV71的病毒复制和组装，EV71基因组上m5C修饰位点突变后显著减弱了病毒复制。同时，EV71 RNA的m5C修饰增强了病毒蛋白的翻译，促进了病毒RNA的稳定性。此外，NSUN2与EV71 VP1互作抑制其K48泛素化促进VP1稳定性。在AG6乳鼠中，与WT EV71相比，m5C修饰缺陷毒株的致病性显着减弱。综上所述，NSUN2和m5C修饰在EV71生命周期中发挥着多重作用。我们解析了NSUN2介导的调控EV71 复制的不同途径，证实了NSUN2催化的EV71 RNA上的m5C修饰对病毒复制和致病性非常重要。
- NSUN2
- , m5C
- , EV71
- , 泛素化
- , 致病性

NSUN2 mediates distinct pathways to regulate enterovirus 71 replication

Lishi Liu ^a,b ,
Zhen Chen ^a ,
Kui Zhang ^a,b ,
Haojie Hao ^a ,
Li Ma ^a,b ,
Haizhou Liu ^a ,
Baocheng Yu ^a,b ,
Shuang Ding ^a ,
Xueyan Zhang ^a ,
Miao Zhu ^a,b ,
Xiang Guo ^a,b ,
Yi Liu ^c ,
Haibin Liu ^a ,
Fang Huang ^c,, ,
Ke Peng ^a,c,, ,
Wuxiang Guan ^a,c,,

a. Center for Emerging Infectious Diseases, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan, Hubei, 430071, China;
b. University of Chinese Academy of Sciences, Beijing, 100049, China;
c. Hubei Jiangxia Laboratory, Wuhan, Hubei, 430200, China

Corresponding author: Fang Huang, huangf@wh.iov.cn Ke Peng, pengke@wh.iov.cn Wuxiang Guan, guanwx@wh.iov.cn
Received Date: 03 March 2024
Accepted Date: 03 April 2024

Abstract

Increasing evidences suggest that the methyltransferase NSUN2 catalyzes 5-methylcytosine (m5C) modifications on viral RNAs, which are essential for the replication of various viruses. Despite the function of m5C deposition is well characterized, other potential roles of NSUN2 in regulating viral replication remain largely unknown. In this study, the m5C modified residues catalyzed by NSUN2 on enterovirus 71 (EV71) RNAs were mapped. NSUN2, along with m5C modifications, played multiple roles during the EV71 life cycle. Functional m5C modified nucleotides increased the translational efficiency and stability of EV71 RNAs. Additionally, NSUN2 was found to target the viral protein VP1 for binding and promote its stability by inhibiting the ubiquitination. Furthermore, both viral replication and pathogenicity in mice were largely attenuated when functional m5C residues were mutated. Taken together, this study characterizes distinct pathways mediated by NSUN2 in regulating EV71 replication, and highlights the importance of its catalyzed m5C modifications on EV71 RNAs for the viral replication and pathogenicity.
- NSUN2
- , 5-methylcytosine
- , Enterovirus 71
- , Ubiquitination
- , Pathogenicity

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10.1016j.virs.2024.05.002-ESM.docx