Differential requirements for COPI coats in formation of replication complexes among three genera of Picornaviridae
Differential requirements for COPI coats in formation of replication complexes among three genera of Picornaviridae
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DOI：
10.1128/jvi.76.21.11113-11122.2002
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发表时间：
2002-11-01
影响因子：
5.4
通讯作者：
Anderson, DA
中科院分区：
文献类型：
--
作者：
Gazina, EV;Mackenzie, JM;Anderson, DA
小核糖核酸病毒RNA复制需要复制复合物（RC）的形成。由与病毒非结构蛋白和RNA相关的病毒诱导的囊泡组成。布雷菲德菌素A（BFA）已被证明能强烈抑制脊髓灰质炎病毒的RNA复制，但不能抑制脑心肌炎病毒（EMCV）。在这里，我们证明了双埃柯病毒1（ParV 1）的复制部分耐BFA，而埃可病毒11（EV 11）的复制被强烈抑制。由于BFA抑制内质网（ER）-高尔基体转运中的COPI依赖步骤，我们测试了一个假设，即不同的小核糖核酸病毒在其RC的形成中可能对COPI有不同的要求。使用免疫荧光和冷冻免疫电子显微镜，我们研究了COPI组分β-COP与EMCV、ParV 1和EV 11的RC的相关性，EMCV RC不含β-COP。相比之下，β-COP似乎特异性地分布于EV 11的RC。在ParV 1感染的细胞中，β-COP大部分分散在整个细胞质中，其中一些存在于RC中。这些结果表明，COPI参与各种小RNA病毒RC的形成存在差异，对应于它们对BFA的不同敏感性。EMCV RC可能在囊泡从ER出芽后立即形成，在COPI与膜结合之前。ParV 1 RC由含COPI的膜形成，但COPI不太可能直接参与其形成，而EV 11 RC的形成似乎取决于COPI与膜的结合。
Picornavirus RNA replication requires the formation of replication complexes (RCs). consisting of virus-induced vesicles associated with viral nonstructural proteins and RNA. Brefeldin A (BFA) has been shown to strongly inhibit RNA replication of poliovirus but not of encephalomyocarditis virus (EMCV). Here, we demonstrate that the replication of parechovirus 1 (ParV1) is partly resistant to BFA, whereas echovirus 11 (EV11) replication is strongly inhibited. Since BFA inhibits COPI-dependent steps in endoplasmic reticulum (ER)-Golgi transport, we tested a hypothesis that different picornaviruses may have differential requirements for COPI in the formation of their RCs. Using immunofluorescence and cryo-immunoelectron microscopy we examined the association of a COPI component, beta-COP, with the RCs of EMCV, ParV1, and EV11 EMCV RCs did not contain beta-COP. In contrast, beta-COP appeared to be specifically distributed to the RCs of EV11 In ParV1-infected cells beta-COP was largely dispersed throughout the cytoplasm, with some being present in the RCs. These results suggest that there are differences in the involvement of COPI in the formation of the RCs of various picornaviruses, corresponding to their differential sensitivity to BFA. EMCV RCs are likely to be formed immediately after vesicle budding from the ER, prior to COPI association with membranes. ParV1 RCs are formed from COPI-containing membranes but COPI is unlikely to be directly involved in their formation, whereas formation of EV11 RCs appears to be dependent on COPI association with membranes.