Functional microRNA screen uncovers O-linked N-acetylglucosamine transferase as a host factor modulating hepatitis C virus morphogenesis and infectivity.
OBJECTIVE
Infection of human hepatocytes by the hepatitis C virus (HCV) is a multistep process involving both viral and host factors. microRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally regulate gene expression. Given that miRNAs were indicated to regulate between 30% and 75% of all human genes, we aimed to investigate the functional and regulatory role of miRNAs for the HCV life cycle.


DESIGN
To systematically reveal human miRNAs affecting the HCV life cycle, we performed a two-step functional high-throughput miRNA mimic screen in Huh7.5.1 cells infected with recombinant cell culture-derived HCV. miRNA targeting was then assessed using a combination of computational and functional approaches.


RESULTS
We uncovered miR-501-3p and miR-619-3p as novel modulators of HCV assembly/release. We discovered that these miRNAs regulate O-linked N-acetylglucosamine (O-GlcNAc) transferase (OGT) protein expression and identified OGT and O-GlcNAcylation as regulators of HCV morphogenesis and infectivity. Furthermore, increased OGT expression in patient-derived liver tissue was associated with HCV-induced liver disease and cancer.


CONCLUSION
miR-501-3p and miR-619-3p and their target OGT are previously undiscovered regulatory host factors for HCV assembly and infectivity. In addition to its effect on HCV morphogenesis, OGT may play a role in HCV-induced liver disease and hepatocarcinogenesis.
['Herzog|Katharina|K|', 'Bandiera|Simonetta|S|', 'Pernot|Sophie|S|', 'Fauvelle|Catherine|C|', 'Jühling|Frank|F|', 'Weiss|Amélie|A|', 'Bull|Anne|A|', 'Durand|Sarah C|SC|', 'Chane-Woon-Ming|Béatrice|B|', 'Pfeffer|Sébastien|S|', 'Mercey|Marion|M|', 'Lerat|Hervé|H|', 'Meunier|Jean-Christophe|JC|', 'Raffelsberger|Wolfgang|W|', 'Brino|Laurent|L|', 'Baumert|Thomas F|TF|0000-0002-9022-5611', 'Zeisel|Mirjam B|MB|0000-0003-1606-0131']
D005786:Gene Expression Regulation D055785:Gene Knockdown Techniques D055106:Genome-Wide Association Study D016174:Hepacivirus D019698:Hepatitis C, Chronic D022781:Hepatocytes D054884:Host-Pathogen Interactions D006801:Humans D008018:Life Cycle Stages D035683:MicroRNAs D009024:Morphogenesis D017351:N-Acetylglucosaminyltransferases D015854:Up-Regulation D014774:Virulence
hepatitis C hepatocyte HCV molecular mechanisms