Coronavirus replication complex formation utilizes components of cellular autophagy.
Coronavirus replication complex formation utilizes components of cellular autophagy.
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DOI：
10.1074/jbc.m306124200
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发表时间：
2004-03-12
期刊：
影响因子：
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通讯作者：
Denison MR
中科院分区：
文献类型：
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作者：
Prentice E;Jerome WG;Yoshimori T;Mizushima N;Denison MR
冠状病毒小鼠肝炎病毒（MHV）在宿主细胞质中的双膜囊泡（dmv）上进行RNA复制。然而，这些dmv形成的机制尚未确定。利用遗传、生化和细胞成像方法，研究了自噬在DMV形成和MHV复制中的作用。结果表明，复制复合体与自噬蛋白、微管相关蛋白轻链3和Apg12共定位。MHV感染通过抵抗3-甲基腺嘌呤抑制的机制诱导自噬。在自噬敲除、APG5 - / -和胚胎干细胞系中，MHV复制受损，但在APG5 - / -细胞中表达APG5可以恢复MHV复制的野生型水平。在mhv感染的APG5 - / -细胞中，未检测到dmv；相反，粗糙的内质网明显肿胀。本研究结果表明，双膜结合MHV复制复合体的形成需要自噬，DMV的形成显著提高了复制效率。此外，粗内质网被认为是复制复合体膜的可能来源。
The coronavirus mouse hepatitis virus (MHV) performs RNA replication on double membrane vesicles (DMVs) in the cytoplasm of the host cell. However, the mechanism by which these DMVs form has not been determined. Using genetic, biochemical, and cell imaging approaches, the role of autophagy in DMV formation and MHV replication was investigated. The results demonstrated that replication complexes co-localize with the autophagy proteins, microtubule-associated protein light-chain 3 and Apg12. MHV infection induces autophagy by a mechanism that is resistant to 3-methyladenine inhibition. MHV replication is impaired in autophagy knockout, APG5–/–, embryonic stem cell lines, but wild-type levels of MHV replication are restored by expression of Apg5 in the APG5–/–cells. In MHV-infected APG5–/–cells, DMVs were not detected; rather, the rough endoplasmic reticulum was dramatically swollen. The results of this study suggest that autophagy is required for formation of double membrane-bound MHV replication complexes and that DMV formation significantly enhances the efficiency of replication. Furthermore, the rough endoplasmic reticulum is implicated as the possible source of membranes for replication complexes.