Biogenesis and molecular characteristics of serum hepatitis B virus RNA

• DOI: 10.1371/journal.ppat.1008945
• Publication Date: 2020-10-20T17:23:18Z
• Authors: Sheng Shen; Zhanglian Xie; Dawei Cai; Xiaoyang Yu; Hu Zhang; Elena S. Kim; Bin Zhou; Jinlin Hou; Xiaoyong Zhang; Qi Huang; Jian Sun; Haitao Guo
• Keywords: 0301 basic medicine; Hepatitis B virus; QH301-705.5; Virus Assembly; DNA; Reverse Transcription; RC581-607; Virus Replication; 3. Good health; 03 medical and health sciences; Capsid; DNA, Viral; Hepatocytes; RNA; Humans; RNA, Viral; Capsid Proteins; Immunologic diseases. Allergy; Biology (General); Research Article

HBV is an enveloped DNA virus that replicates its genome via reverse transcription of a pregenomic (pg) RNA intermediate in hepatocytes. Interestingly, can be detected virus-like particles chronic hepatitis B (CHB) patient serum and has been utilized as biomarker for intrahepatic cccDNA activity treated patients. However, the biogenesis molecular characteristics remain to fully defined. In this study, we found encapsidated predominately consists pgRNA, which are detergent- ribonuclease-resistant. Through blocking replication without affecting pgRNA encapsidation by using priming-defective mutant Y63D or 3TC treatment, demonstrated cell culture supernatant contains large amount pgRNA-containing nonenveloped capsids minor population virions. The formation pgRNA-virion requires both capsid assembly viral envelope proteins, inhibited modulators envelope–knockout mutant, respectively. Furthermore, utilizes multivesicular body pathway egress, similar way DNA-virion morphogenesis. Northern blotting, RT-PCR, 3' RACE assays revealed serum/supernatant mainly spliced devoid 3'-terminal sequences. collected from cells with reversible priming inhibitor L-FMAU was unable establish infection HepG2-NTCP cells. summary, secreted noninfectious virion-like particle poly(A)-free pgRNA. Our study will shed light on biology life cycle, aid development novel CHB diagnosis treatment prognosis.