Epitranscriptomic cytidine methylation of the hepatitis B viral RNA is essential for viral reverse transcription and particle production

• 成果类型: Article
• 署名作者: Su, Pei-Yi (Alma); Chang, Chih- Hsu; Yen, Shin-Chwen Bruce; Wu, Hsiu- Yi; Tung, Wan- Ju; Hu, Yu- Pei; Chen, Yen-Yu Ian; Lin, Miao- Hsia; Shih, Chiaho; Chen, Pei- Jer; Tsai, Kevin
• 署名单位: Academia Sinica - Taiwan; National Yang Ming Chiao Tung University; Academia Sinica - Taiwan; Academia Sinica - Taiwan; National Taiwan University; China Medical University Taiwan; National Taiwan University; National Taiwan University; National Taiwan University; National Taiwan University Hospital
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-14423
• DOI: 10.1073/pnas.2400378121
• 发表日期: 2024-06-11
• 关键词: messenger-rna; wide identification; packaging signal; stem-loop; 5-methylcytosine; protein; cells; dna; encapsidation; inhibitors

摘要

Epitranscriptomic RNA modifications have emerged as important regulators of the fate and function of viral RNAs. One prominent modification, the cytidine methylation 5 - methylcytidine (m 5 C), is found on the RNA of HIV - 1, where m 5 C enhances the translation of HIV - 1 RNA. However, whether m 5 C functionally enhances the RNA of other pathogenic viruses remains elusive. Here, we surveyed a panel of commonly found RNA modifications on the RNA of hepatitis B virus (HBV) and found that HBV RNA is enriched with m 5 C as well as ten other modifications, at stoichiometries much higher than host messenger RNA (mRNA). Intriguingly, m 5 C is mostly found on the epsilon hairpin, an RNA element required for viral RNA encapsidation and reverse transcription, with these m 5 C mainly deposited by the cellular methyltransferase NSUN2. Loss of m 5 C from HBV RNA due to NSUN2 depletion resulted in a partial decrease in viral core protein (HBc) production, accompanied by a near - complete loss of the reverse transcribed viral DNA. Similarly, mutations introduced to remove the methylated cytidines resulted in a loss of HBc production and reverse transcription. Furthermore, pharmacological disruption of m 5 C deposition led to a significant decrease in HBV replication. Thus, our data indicate m 5 C methylations as a critical mediator of the epsilon elements' function in HBV virion production and reverse transcription, suggesting the therapeutic potential of targeting the m 5 C methyltransfer process on HBV epsilon as an antiviral strategy.