Journal Article

Microrna-130a Downregulates HCV Replication through an atg5-Dependent Autophagy Pathway

Li, Yujia, Sheng, Qiuju, Chen, Dong, Chung, Raymond T., Cheng, Zhimeng, Xu, Min, Kruger, Annie J., Li, Shuang, Li, Shilin, Liu, Xiao, Chen, Limin, Kaj, Batul, Lin, Wenyu, Duan, Xiaoqiong, Liao, Xinzhong, Li, Wenting, Holmes, Jacinta A., Shao, Tuo, Schaefer, Esperance A., Yang, Chunhui, Ye, Haiyan

2019

Overview

We previously identified that miR-130a downregulates HCV replication through two independent pathways: restoration of host immune responses and regulation of pyruvate metabolism. In this study, we further sought to explore host antiviral target genes regulated by miR-130a. We performed a RT² Profiler™ PCR array to identify the host antiviral genes regulated by miR-130a. The putative binding sites between miR-130a and its downregulated genes were predicted by miRanda. miR-130a and predicted target genes were over-expressed or knocked down by siRNA or CRISPR/Cas9 gRNA. Selected gene mRNAs and their proteins, together with HCV replication in JFH1 HCV-infected Huh7.5.1 cells were monitored by qRT-PCR and Western blot. We identified 32 genes that were significantly differentially expressed more than 1.5-fold following miR-130a overexpression, 28 of which were upregulated and 4 downregulated. We found that ATG5, a target gene for miR-130a, significantly upregulated HCV replication and downregulated interferon stimulated gene expression. miR-130a downregulated ATG5 expression and its conjugation complex with ATG12. ATG5 and ATG5-ATG12 complex affected interferon stimulated gene (ISG) such as MX1 and OAS3 expression and subsequently HCV replication. We concluded that miR-130a regulates host antiviral response and HCV replication through targeting ATG5 via the ATG5-dependent autophagy pathway.

• Publisher: MDPI AG,MDPI
• Subject: Antibodies / Autophagy / autophagy-related genes 5 (ATG5), ATG12 / Autophagy-Related Protein 12 - metabolism / Autophagy-Related Protein 5 - metabolism / Binding sites / Cell Line / Cloning / CRISPR / Gene expression / Gene Expression Regulation / Genes, Regulator / gRNA / Hepacivirus - physiology / Hepatitis C / Hepatitis C - virology / hepatitis C virus (HCV), interferon stimulated gene (ISG) / Humans / Immune response / Infections / Interferon / MicroRNAs / MicroRNAs - physiology / miR-130a / miRNA / Myxovirus resistance proteins / Phagocytosis / Plasmids / Proteins / Pyruvic acid / Reagents / Replication / siRNA / Viral infections / Virus Replication / Viruses