The PRMT5/WDR77 complex restricts hepatitis E virus replication.
The PRMT5/WDR77 complex restricts hepatitis E virus replication.
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DOI：
10.1371/journal.ppat.1011434
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发表时间：
2023-06
期刊：
影响因子：
6.7
通讯作者：
中科院分区：
文献类型：
--
作者：
戊型肝炎病毒（Hepatitis E virus，HEV）是世界范围内急性肝炎的主要病原体之一。HEV的复制机制，尤其是宿主因素对HEV复制的调控机制尚不清楚。本研究利用HEV ORF 1反式互补细胞培养系统和HEV复制子系统，结合细胞培养中氨基酸稳定同位素标记（SILAC）和质谱（MS）技术，对HEV复制的宿主调控因子进行了研究。在这两种细胞培养模型中，我们鉴定了多种与HEV ORF 1蛋白相关的宿主因子，这些宿主因子可能负责病毒基因组RNA的复制。值得注意的是，蛋白质精氨酸甲基转移酶5（PRMT 5）/WDR 77复合物在两种细胞培养模型中被鉴定为最高命中。此外，我们证明PRMT 5和WDR 77可以特异性地抑制HEV复制，但不能抑制其他病毒如HCV或SARS-CoV-2，并且这种抑制在不同的HEV毒株和基因型中是保守的。PRMT 5/WDR 77可催化ORF 1的R458甲基化，从而削弱其复制酶活性，而ORF 1中携带R458 K突变的病毒相应地解除PRMT 5/WDR 77的限制。总之，我们的研究促进了对病毒感染的更全面的了解，但也为干预提供了治疗靶点。HEV是一种被低估的病原体，每年在全球范围内造成约2000万例感染，并导致约70，000人死亡。没有直接作用于HEV治疗的抗病毒药物。虽然关于HEV复制的研究已经取得了很大的进展，但是宿主因子对HEV复制的调控作用还没有得到充分的研究。本文利用ORF 1反式互补系统和在ORF 1中携带HA-Flag标签的HEV复制子，系统地分析了病毒复制条件下HEV复制复合体中的宿主因素。蛋白精氨酸甲基转移酶5（PRMT 5）/WDR 77复合物存在于HEV复制复合物中，对HEV不同毒株间的感染有抑制作用，但对HCV和SARS-CoV-2感染无抑制作用。PRMT 5/WDR 77复合物能够在负责HEV复制的第458位精氨酸处甲基化HEV ORF 1，从而抑制HEV复制。ORF 1区R458 K置换后的HEV能逃避PRMT 5/WDR 77的限制，复制能力增强。这项工作不仅为HEV复制和病毒-宿主相互作用提供了新的见解，而且为针对HEV感染的抗病毒策略提供了信息。
Hepatitis E virus (HEV) is one of the main pathogenic agents of acute hepatitis in the world. The mechanism of HEV replication, especially host factors governing HEV replication is still not clear. Here, using HEV ORF1 trans-complementation cell culture system and HEV replicon system, combining with stable isotope labelling with amino acids in cell culture (SILAC) and mass spectrometry (MS), we aimed to identify the host factors regulating HEV replication. We identified a diversity of host factors associated with HEV ORF1 protein, which were putatively responsible for viral genomic RNA replication, in these two cell culture models. Of note, the protein arginine methyltransferase 5 (PRMT5)/WDR77 complex was identified in both cell culture models as the top hit. Furthermore, we demonstrated that PRMT5 and WDR77 can specifically inhibit HEV replication, but not other viruses such as HCV or SARS-CoV-2, and this inhibition is conserved among different HEV strains and genotypes. Mechanistically, PRMT5/WDR77 can catalyse methylation of ORF1 on its R458, impairing its replicase activity, and virus bearing R458K mutation in ORF1 relieves the restriction of PRMT5/WDR77 accordingly. Taken together, our study promotes more comprehensive understanding of viral infections but also provides therapeutic targets for intervention. HEV is an underestimated pathogen, causing about 20 million infections worldwide every year and leading to approximately 70,000 deaths. There are no direct-acting antivirals for HEV treatment. Although significant progress has been made about HEV replication, but host factors regulating HEV replication have not been fully characterized. Here, using ORF1 trans-complementation system and HEV replicon harbouring HA-Flag tag in ORF1, we systematically analyse host factors in HEV replication complex in viral replication condition. Specifically, protein arginine methyltransferase 5 (PRMT5)/WDR77 complex was found to be in HEV replication complex and play an inhibitory role in HEV infection among different HEV strain, but not in HCV and SARS-CoV-2 infection. In details, PRMT5/WDR77 complex is able to methylate HEV ORF1 at 458th arginine, which is responsible for HEV replication, to repress HEV replication. Consistently, HEV with R458K substitution in ORF1 escape the restriction by PRMT5/WDR77, with enhanced replication capability. This work not only provides novel insights into HEV replication and viral-host interaction, but also informs the antiviral strategy against HEV infection.
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SM级核糖核蛋白的生物发生需要两个不同的精氨酸甲基转移酶。
影响因子：
7.8
作者：
Gonsalvez, Graydon B.;Tian, Liping;Ospina, Jason K.;Boisvert, Francois-Michel;Lamond, Angus I.;Matera, A. Gregory
通讯作者：
Matera, A. Gregory
DOI：
10.1007/978-3-319-46503-6_7
发表时间：
2017-01-01
期刊：
MACROMOLECULAR PROTEIN COMPLEXES: STRUCTURE AND FUNCTION
影响因子：
--
作者：
Antonysamy, Stephen
通讯作者：
Antonysamy, Stephen
影响因子：
64.5
作者：
Hsu PD;Lander ES;Zhang F
通讯作者：
Zhang F
影响因子：
6.4
作者：
Ding Q;Nimgaonkar I;Archer NF;Bram Y;Heller B;Schwartz RE;Ploss A
通讯作者：
Ploss A
影响因子：
5.4
作者：
Benhenda, Shirine;Ducroux, Aurelie;Neuveut, Christine
通讯作者：
Neuveut, Christine