Quantitative Proteomics Identifies Serum Response Factor Binding Protein 1 as a Host Factor for Hepatitis C Virus Entry

Hepatitis C virus (HCV) enters human hepatocytes through a multistep mechanism involving, among other host proteins, the virus receptor CD81. How CD81 governs HCV entry is poorly characterized, and CD81 protein interactions after virus binding remain elusive. We have developed a quantitative proteomics protocol to identify HCV-triggered CD81 interactions and found 26 dynamic binding partners. At least six of these proteins promote HCV infection, as indicated by RNAi. We further characterized serum response factor binding protein 1 (SRFBP1), which is recruited to CD81 during HCV uptake and supports HCV infection in hepatoma cells and primary human hepatocytes. SRFBP1 facilitates host cell penetration by all seven HCV genotypes, but not of vesicular stomatitis virus and human coronavirus. Thus, SRFBP1 is an HCV-specific, pan-genotypic host entry factor. These results demonstrate the use of quantitative proteomics to elucidate pathogen entry and underscore the importance of host protein-protein interactions during HCV invasion. [Display omitted] •Hepatitis C virus binding alters host protein interactions with the receptor CD81•Six out of 26 virus-dependent CD81-interacting proteins promote virus entry•SRFBP1 binds CD81 and aids infection of all HCV, but not VSV, genotypes•SRFBP1 is membrane-associated and required for HCV entry Hepatitis C virus (HCV) enters human hepatocytes through a multistep mechanism. Gerold et al. apply quantitative proteomics to define the protein network responsible for HCV entry and identify SRFBP1 as a partner for the virus receptor CD81.