The autophagy elongation complex (ATG5-12/16L1) positively regulates HCV replication and is required for wild-type membranous web formation.
The autophagy elongation complex (ATG5-12/16L1) positively regulates HCV replication and is required for wild-type membranous web formation.
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DOI：
10.1038/srep40351
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发表时间：
2017-01-09
影响因子：
4.6
通讯作者：
Labonté P
中科院分区：
文献类型：
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作者：
Fahmy AM;Labonté P
丙型肝炎病毒（HCV）感染诱导细胞内膜重排，从而形成膜网（MW），其中HCV复制和组装发生。HCV诱导的MW主要由双膜囊泡（DMV）和多膜囊泡组成。自噬机制被认为参与了这种囊泡的形成。然而，没有发现明确的证据将自噬与这些DMV的形成联系起来。在这项研究中，我们评估了自噬延伸复合物（ATG 5 -12/16 L1）在HCV复制和MW形成中的作用。使用ATG 12的显性阴性形式和siRNA方法，我们证明了ATG 5 -12缀合物而不是LC 3-II形成对于有效的病毒复制是至关重要的。此外，HCV MW的纯化揭示了ATG 5 -12和ATG 16 L1沿着HCV非结构蛋白的存在。有趣的是，LC 3没有沿着延伸复合物被募集到病毒复制的位点。最后，延长复合物的抑制，但不是LC 3，大大削弱了野生型MW表型的形成。据我们所知，这项研究提供了自噬蛋白参与野生型MW形成的第一个证据。
Hepatitis C virus (HCV) infection induces intracellular membrane rearrangements, thus forming a membranous web (MW) in which HCV replication and assembly occur. The HCV-induced MW is primarily composed of double membrane vesicles (DMVs) transfused by multi-membrane vesicles. The autophagy machinery has been proposed to participate in the formation of such vesicles. However, no clear evidence has been found linking autophagy to the formation of these DMVs. In this study, we evaluated the role of the autophagy elongation complex (ATG5-12/16L1) in HCV replication and MW formation. Using a dominant negative form of ATG12 and an siRNA approach, we demonstrated that the ATG5-12 conjugate, but not LC3-II formation, is crucial for efficient viral replication. Furthermore, purification of HCV MW revealed the presence of ATG5-12 and ATG16L1 along with HCV nonstructural proteins. Interestingly, LC3 was not recruited along with the elongation complex to the site of viral replication. Finally, inhibition of the elongation complex, but not LC3, greatly impaired the formation of the wild-type MW phenotype. To our knowledge, this study provides the first evidence of the involvement of autophagy proteins in the formation of wild-type MWs.