Epidermal growth factor receptor is a host-entry cofactor triggering hepatitis B virus internalization

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 116; no. 17; pp. 8487 - 8492
• Main Authors: Iwamoto, Masashi, Saso, Wakana, Sugiyama, Ryuichi, Ishii, Koji, Ohki, Mio, Nagamori, Shushi, Suzuki, Ryosuke, Aizaki, Hideki, Ryo, Akihide, Yun, Ji-Hye, Park, Sam-Yong, Ohtani, Naoko, Muramatsu, Masamichi, Iwami, Shingo, Tanaka, Yasuhito, Sureau, Camille, Wakita, Takaji, Watashi, Koichi
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 23.04.2019
• Subjects: Bile; Biological Sciences; Cell culture; Cell membranes; Cell surface; Deactivation; Epidermal growth factor; Epidermal growth factor receptors; ErbB Receptors - genetics; ErbB Receptors - metabolism; Fluorescence; Growth factors; Hep G2 Cells; Hepatitis; Hepatitis B; Hepatitis B virus - pathogenicity; Hepatitis B virus - physiology; Host-Pathogen Interactions - genetics; Host-Pathogen Interactions - physiology; Humans; Immunoprecipitation; Inactivation; Infections; Internalization; Life Sciences; Microbiology and Parasitology; Molecular interactions; Mutation; Organic Anion Transporters, Sodium-Dependent - genetics; Organic Anion Transporters, Sodium-Dependent - metabolism; Peptides; Pharmacology; Point mutation; Sodium; Symporters - genetics; Symporters - metabolism; Viral infections; Virions; Virology; Virus Internalization; Viruses; entry; HBV; EGFR; NTCP; transporter
• ISSN: 0027-8424; 1091-6490; 1091-6490
• DOI: 10.1073/pnas.1811064116

Sodium taurocholate cotransporting polypeptide (NTCP) is a host cell receptor required for hepatitis B virus (HBV) entry. However, the susceptibility of NTCP-expressing cells to HBV is diverse depending on the culture condition. Stimulation with epidermal growth factor (EGF) was found to potentiate cell susceptibility to HBV infection. Here, we show that EGF receptor (EGFR) plays a critical role in HBV virion internalization. In EGFR-knockdown cells, HBV or its preS1-specific fluorescence peptide attached to the cell surface, but its internalization was attenuated. PreS1 internalization and HBV infection could be rescued by complementation with functional EGFR. Interestingly, the HBV/preS1–NTCP complex at the cell surface was internalized concomitant with the endocytotic relocalization of EGFR. Molecular interaction between NTCP and EGFR was documented by immunoprecipitation assay. Upon dissociation from functional EGFR, NTCP no longer functioned to support viral infection, as demonstrated by either (i) the introduction of NTCP pointmutation that disrupted its interaction with EGFR, (ii) the detrimental effect of decoy peptide interrupting the NTCP–EGFR interaction, or (iii) the pharmacological inactivation of EGFR. Together, these data support the crucial role of EGFR in mediating HBV–NTCP internalization into susceptible cells. EGFR thus provides a yet unidentified missing link from the cell-surface HBV–NTCP attachment to the viral invasion beyond the host cell membrane