Characterization of CD81 receptor interactors in hepatitis C virus and Plasmodium liver cell entry.
Final Report Abstract
The tetraspanin CD81 is a plasma membrane protein that interacts with a variety of proteins and lipids. In healthy cells, CD81 plays an important role during cell development, activation by external stimuli and motility. In addition, two pathogens, hepatitis C virus (HCV) and the malaria parasite Plasmodium, hijack CD81 to enter liver cells. Previously it remained largely elusive how CD81 guides the entry process of both pathogens and whether similar mechanisms are used by the virus and the parasite. In this study, we mapped CD81 interacting proteins in human liver derived cell lines and in primary human hepatocytes using quantitative proteomics. In total, we identified 42 CD81 interacting proteins including known HCV host factors (e.g. EGFR). In addition, we discovered two novel CD81 interaction partners, namely calpain-5 (CAPN5) and Casitas B-lineage lymphoma proto-oncogene B (CBLB), which facilitate HCV infection. Knocking out CAPN5 and CBLB in hepatoma cells by CRISPR/Cas9 technology led to reduced HCV susceptibility for all seven HCV genotypes tested. In contrast, other enveloped RNA viruses such as human coronavirus and vesicular stomatitis virus did not depend on CAPN5 and CBLB, suggesting that both proteins are HCV-specific and pangenotypic host factors. Moreover, Plasmodium did not require CAPN5 or CBLB for invasion into liver cells. Instead, two other CD81 interacting proteins, the EGFR adaptor protein GRB2 and the ubiquitin ligase CBL aided Plasmodium invasion. During HCV infection, CAPN5 and CBLB functioned independently of their enzymatic activity or signaling function suggesting a scaffolding function. By dissecting the entry steps of the virus into liver cells, we could show that CAPN5 and CBLB facilitate a post-binding and pre-fusion step of HCV entry, i.e., endocytosis or endosomal trafficking. In sum, we mapped CD81 protein interactions in human liver cells and showed that HCV and Plasmodium differently use CD81 interaction partners for liver cell entry.
Publications
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2020. Cholesterol sensing by CD81 is important for hepatitis C virus entry. J. Biol. Chem. 295, 16931–16948
Palor, M., Stejskal, L., Mandal, P., Lenman, A., Alberione, M.P., Kirui, J., Moeller, R., Ebner, S., Meissner, F., Gerold, G., Shepherd, A.J., Grove, J.
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2017. Protein Interactions during the Flavivirus and Hepacivirus Life Cycle. Mol. Cell Proteomics 16, S75–S91
Gerold, G., Bruening, J., Weigel, B., Pietschmann, T.
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2018. CD81 Receptor Regions outside the Large Extracellular Loop Determine Hepatitis C Virus Entry into Hepatoma Cells. Viruses 10
Banse, P., Moeller, R., Bruening, J., Lasswitz, L., Kahl, S., Khan, A.G., Marcotrigiano, J., Pietschmann, T., Gerold, G.
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2018. Hepatitis C virus enters liver cells using the CD81 receptor complex proteins calpain-5 and CBLB. PLoS Pathog. 14, e1007111
Bruening, J., Lasswitz, L., Banse, P., Kahl, S., Marinach, C., Vondran, F.W., Kaderali, L., Silvie, O., Pietschmann, T., Meissner, F., Gerold, G.
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2020. Hepatitis C virus entry: protein interactions and fusion determinants governing productive hepatocyte invasion. Cold Spring Harb. Perspect. Med. 10
Gerold, G., Moeller, R., Pietschmann, T.
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2020. Single-nucleotide variants in human CD81 influence hepatitis C virus infection of hepatoma cells. Med Microbiol Immunol 209, 499–514
Alberione, M.P., Moeller, R., Kirui, J., Ginkel, C., Doepke, M., Ströh, L.J., Machtens, J.-P., Pietschmann, T., Gerold, G.
