Knockdown of autophagy-related gene decreases the production of infectious hepatitis C virus particles
Knockdown of autophagy-related gene decreases the production of infectious hepatitis C virus particles
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DOI：
10.4161/auto.5.7.9243
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发表时间：
2009-10-01
期刊：
影响因子：
13.3
通讯作者：
Hanada, Kentaro
中科院分区：
文献类型：
--
作者：
Tanida, Isei;Fukasawa, Masayoshi;Hanada, Kentaro
丙型肝炎病毒是一种正链RNA病毒，属于黄病毒科。当丙型肝炎病毒JFH1RNA和丙型肝炎病毒亚基因组复制子被导入Huh7.5细胞时，ATG7-基因敲除减少了丙型肝炎病毒复制子RNA的量。然而，当感染性的原始丙型肝炎病毒颗粒直接感染Huh7.5.1细胞时，ATG7基因敲除是否会减少细胞内丙型肝炎病毒相关蛋白、丙型肝炎病毒mRNA和感染性丙型肝炎病毒颗粒的产生仍不清楚。用RNA干扰技术下调感染丙型肝炎病毒的Huh7.5.1细胞的ATG7蛋白后，细胞内丙型肝炎病毒核心蛋白、NS3、NS5A蛋白、丙型肝炎病毒信使核糖核酸和分泌白蛋白水平与对照组相比无明显变化。然而，ATG7基因敲除后，在培养液中释放的感染性丙型肝炎病毒颗粒水平降低。当Beclin I被RNA干扰击倒时，也得到了类似的结果。此外，还研究了内源性LC3-斑点蛋白与丙型肝炎病毒核心蛋白、HS5A蛋白和脂滴的共存情况。但很少有内源性的Lc3点与丙型肝炎病毒核心蛋白、NS5A蛋白或脂滴共存。这些结果表明，在一个原始的丙型肝炎病毒颗粒感染系统中，自噬有助于有效地产生丙型肝炎病毒颗粒，但对细胞内丙型肝炎病毒相关蛋白、丙型肝炎病毒信使核糖核酸和分泌途径的产生几乎没有贡献。
Hepatitis C virus (HCV) is a positive-strand RNA virus, and classified within the Flaviridae family. Atg7-knockdown decreases the amount of HCV replicon RNA, when HCV JFH1 RNA and HCV subgenomic replicon are transfected into Huh7.5 cells. However, when infectious naive HCV particles are directly infected into Huh7.5.1 cells, it is still unclear whether Atg7-knockdown decreases the production of intracellular HCV-related proteins, HCV mRNA and infectious HCV particles. When Atg7 protein in HCV-infected Huh7.5.1 cells was knocked down by RNA-interference, the levels of intracellular HCV core, NS3, NS5A proteins, HCV mRNA and secreted albumin remained unchanged compared with those in the control HCV-infected cells. However, the level of infectious HCV particles released in the medium was decreased by the Atg7-knockdown. Similar results were obtained when Beclin I was knocked down by RNA-interference. The colocalization of endogenous LC3-puncta with HCV core, HS5A proteins and lipid droplets was also investigated. However, little endogenous LC3-puncta colocalized with HCV core, NS5A proteins or lipid droplets. These results suggested that autophagy contributed to the effective production of HCV particles, but little to the intracellular production of HCV-related proteins, HCV mRNA and the secretory pathway, in a naive HCV particles-infection system.