STUB1 regulates antiviral RNAi through inducing ubiquitination and degradation of Dicer and AGO2 in mammals

Shumin Zhang; Xuhua Zhang; Yuanyuan Bie; Jing Kong; An Wang; Yang Qiu; Xi Zhou

doi:10.1016/j.virs.2022.05.001

August 2022

Shumin Zhang, Xuhua Zhang, Yuanyuan Bie, Jing Kong, An Wang, Yang Qiu and Xi Zhou. STUB1 regulates antiviral RNAi through inducing ubiquitination and degradation of Dicer and AGO2 in mammals[J]. Virologica Sinica, 2022, 37(4): 569-580. doi: 10.1016/j.virs.2022.05.001

Citation: Shumin Zhang, Xuhua Zhang, Yuanyuan Bie, Jing Kong, An Wang, Yang Qiu, Xi Zhou. STUB1 regulates antiviral RNAi through inducing ubiquitination and degradation of Dicer and AGO2 in mammals .VIROLOGICA SINICA, 2022, 37(4) : 569-580. http://dx.doi.org/10.1016/j.virs.2022.05.001

STUB1在哺乳动物中通过诱导Dicer和AGO2蛋白的泛素化及降解继而调控抗病毒RNAi

张淑敏 ^a,b ,
张旭华 ^a,b ,
别渊源 ^a,b ,
孔静 ^a,b ,
王岸 ^a,b ,
邱洋 ^a,b,, ,
周溪 ^a,b,,

中国科学院武汉病毒研究所, 中国科学院大学

通讯作者： 邱洋, yangqiu@wh.iov.cn; 周溪, zhouxi@wh.iov.cn
收稿日期： 2022-03-01
录用日期： 2022-04-13

摘要

RNA干扰（RNAi）是真核生物中保守存在的抗病毒免疫机制。然而，哺乳动物中抗病毒RNAi的调控机制还不清楚。在这项研究中，我们发现E3泛素连接酶STUB1是哺乳动物中RNAi通路的一个新的调控因子。我们发现STUB1与AGO2相互作用并使其泛素化，并以伴侣依赖的方式使其降解。STUB1促进了AGO2上K48连接的聚泛素链的形成，并通过泛素-蛋白体系统促进了AGO2的降解。除了AGO2，STUB1还诱导了AGO1、AGO3和AGO4的蛋白降解。进一步发现，STUB1还通过K48连接的聚泛素调节Dicer的泛素化，并诱导Dicer的降解，以及其特殊形式，即在哺乳动物干细胞中表达的抗病毒Dicer（AviDicer）。此外，我们发现STUB1的缺失会上调Dicer和AGO2，从而增强RNAi反应并有效地抑制哺乳动物细胞中的病毒复制。利用肠道病毒71型（EV-A71）的新生小鼠模型，我们证实了敲降STUB1增强了病毒衍生的siRNAs的产生和抗病毒RNAi，在体内引起了对EV-A71的有效抗病毒反应。综上，我们的研究结果发现，STUB1是一种针对RNAi通路的广谱调节因子，靶向Dicer、AviDicer和AGO1-4。此外，STUB1通过调解病毒感染期间Dicer和AGO2的丰度来调节RNAi免疫。我们的研究为哺乳动物中抗病毒RNAi的调控机制提供了新的见解。
- 抗病毒RNAi
- , STUB1
- , AGO2
- , Dicer

STUB1 regulates antiviral RNAi through inducing ubiquitination and degradation of Dicer and AGO2 in mammals

Shumin Zhang ^a,b ,
Xuhua Zhang ^a,b ,
Yuanyuan Bie ^a,b ,
Jing Kong ^a,b ,
An Wang ^a,b ,
Yang Qiu ^a,b,, ,
Xi Zhou ^a,b,,

a State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China;

Corresponding author: Yang Qiu, yangqiu@wh.iov.cn Xi Zhou, zhouxi@wh.iov.cn
Received Date: 01 March 2022
Accepted Date: 13 April 2022

Abstract

RNA interference (RNAi) is an intrinsic antiviral immune mechanism conserved in diverse eukaryotic organisms. However, the mechanism by which antiviral RNAi in mammals is regulated is poorly understood. In this study, we uncovered that the E3 ubiquitin ligase STIP1 homology and U-box-containing protein 1 (STUB1) was a new regulator of the RNAi machinery in mammals. We found that STUB1 interacted with and ubiquitinated AGO2, and targeted it for degradation in a chaperon-dependent manner. STUB1 promoted the formation of Lys48 (K48)-linked polyubiquitin chains on AGO2, and facilitated AGO2 degradation through ubiquitin-proteasome system. In addition to AGO2, STUB1 also induced the protein degradation of AGO1, AGO3 and AGO4. Further investigation revealed that STUB1 also regulated Dicer's ubiquitination via K48-linked polyubiquitin and induced the degradation of Dicer as well as its specialized form, termed antiviral Dicer (aviDicer) that expresses in mammalian stem cells. Moreover, we found that STUB1 deficiency up-regulated Dicer and AGO2, thereby enhancing the RNAi response and efficiently inhibiting viral replication in mammalian cells. Using the newborn mouse model of Enterovirus A71 (EV-A71), we confirmed that STUB1 deficiency enhanced the virus-derived siRNAs production and antiviral RNAi, which elicited a potent antiviral effect against EV-A71 infection in vivo. In summary, our findings uncovered that the E3 ubiquitin ligase STUB1 was a general regulator of the RNAi machinery by targeting Dicer, aviDicer and AGO1–4. Moreover, STUB1 regulated the RNAi response through mediating the abundance of Dicer and AGO2 during viral infection, thereby providing novel insights into the regulation of antiviral RNAi in mammals.
- Antiviral RNAi
- , STIP1 homology and U-box-containing
- , protein 1 (STUB1)
- , Argonaute 2 (AGO2)

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