Hepatitis B virus hijacks TSG101 to facilitate egress via multiple vesicle bodies

Yingcheng Zheng; Mengfei Wang; Sitong Li; Yanan Bu; Zaichao Xu; Guoguo Zhu; Chuanjian Wu; Kaitao Zhao; Aixin Li; Quan Chen; Jingjing Wang; Rong Hua; Yan Teng; Li Zhao; Xiaoming Cheng; Yuchen Xia

doi:10.1371/journal.ppat.1011382

Hepatitis B virus hijacks TSG101 to facilitate egress via multiple vesicle bodies

PLoS Pathog. 2023 May 24;19(5):e1011382. doi: 10.1371/journal.ppat.1011382. eCollection 2023 May.

Authors

Yingcheng Zheng¹, Mengfei Wang¹, Sitong Li¹, Yanan Bu¹, Zaichao Xu¹, Guoguo Zhu², Chuanjian Wu¹, Kaitao Zhao¹, Aixin Li¹, Quan Chen¹, Jingjing Wang¹, Rong Hua¹, Yan Teng¹, Li Zhao¹, Xiaoming Cheng^{1

3

4}, Yuchen Xia¹

Affiliations

¹ State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Hubei Jiangxia Laboratory, Institute of Medical Virology, TaiKang Center for Life and Medical Sciences, TaiKang Medical School, Wuhan University, Wuhan, China.
² Department of Emergency, General Hospital of Central Theater Command of People's Liberation Army of China, Wuhan, China.
³ Wuhan University Center for Pathology and Molecular Diagnostics, Zhongnan Hospital of Wuhan University, Wuhan, China.
⁴ Hubei Clinical Center and Key Laboratory of Intestinal and Colorectal Diseases, Wuhan, China.

Abstract

Hepatitis B virus (HBV) chronically infects 296 million individuals and there is no cure. As an important step of viral life cycle, the mechanisms of HBV egress remain poorly elucidated. With proteomic approach to identify capsid protein (HBc) associated host factors and siRNA screen, we uncovered tumor susceptibility gene 101 (TSG101). Knockdown of TSG101 in HBV-producing cells, HBV-infected cells and HBV transgenic mice suppressed HBV release. Co-immunoprecipitation and site mutagenesis revealed that VFND motif in TSG101 and Lys-96 ubiquitination in HBc were essential for TSG101-HBc interaction. In vitro ubiquitination experiment demonstrated that UbcH6 and NEDD4 were potential E2 ubiquitin-conjugating enzyme and E3 ligase that catalyzed HBc ubiquitination, respectively. PPAY motif in HBc and Cys-867 in NEDD4 were required for HBc ubiquitination, TSG101-HBc interaction and HBV egress. Transmission electron microscopy confirmed that TSG101 or NEDD4 knockdown reduces HBV particles count in multivesicular bodies (MVBs). Our work indicates that TSG101 recognition for NEDD4 ubiquitylated HBc is critical for MVBs mediated HBV egress.

Copyright: © 2023 Zheng et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
DNA-Binding Proteins / genetics
Hepatitis B virus* / genetics
Mice
Mice, Transgenic
Proteomics*
Transcription Factors / genetics

Substances

Tsg101 protein
Transcription Factors
DNA-Binding Proteins

Grants and funding

This work was supported by the National Natural Science Foundation of China (project no. 81971936 to YX), Hubei Province’s Outstanding Medical Academic Leader Program (to YX), Foundation for Innovative Research Groups of the Natural Science Foundation of Hubei (project no. 2020CFA015 to YX), the Fundamental Research Funds for the Central Universities (project no. 2042022kf1215 to XC and 2042021gf0013 to YX) and Basic and Clinical Medical Research Joint Fund of Zhongnan Hospital, Wuhan University (to YX). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.