Quantitative Lipid Droplet Proteome Analysis Identifies Annexin A3 as a Cofactor for HCV Particle Production

Lipid droplets are vital to hepatitis C virus (HCV) infection as the putative sites of virion assembly, but morphogenesis and egress of virions remain ill defined. We performed quantitative lipid droplet proteome analysis of HCV-infected cells to identify co-factors of that process. Our results demonstrate that HCV disconnects lipid droplets from their metabolic function. Annexin A3 (ANXA3), a protein enriched in lipid droplet fractions, strongly impacted HCV replication and was characterized further: ANXA3 is recruited to lipid-rich fractions in HCV-infected cells by the viral core and NS5A proteins. ANXA3 knockdown does not affect HCV RNA replication but severely impairs virion production with lower specific infectivity and higher density of secreted virions. ANXA3 is essential for the interaction of viral envelope E2 with apolipoprotein E (ApoE) and for trafficking, but not lipidation, of ApoE in HCV-infected cells. Thus, we identified ANXA3 as a regulator of HCV maturation and egress. [Display omitted] •Quantitative lipid droplet proteome analysis of HCV-infected cells is presented•ANXA3 is recruited to lipid-rich fractions in HCV-infected cells•ANXA3 functions as a host factor required for efficient HCV particle production•ANXA3 promotes virion maturation by facilitating incorporation of apolipoprotein E Cytoplasmic lipid droplets are vital to hepatitis C virus particle production. Rösch et al. report a quantitative lipid droplet proteome analysis and identify annexin A3 as a regulator of HCV particle maturation and egress.