Identification of functional cis-acting RNA elements in the hepatitis E virus genome required for viral replication


Autoři: Xiaohui Ju aff001;  Guangtao Xiang aff001;  Mingli Gong aff001;  Rui Yang aff002;  Jierui Qin aff001;  Yafei Li aff003;  Yuchen Nan aff003;  Yonglin Yang aff004;  Qiangfeng Cliff Zhang aff002;  Qiang Ding aff001
Působiště autorů: Center for Infectious Disease Research, School of Medicine, Tsinghua University, Beijing, China aff001;  School of Life Sciences, Tsinghua University, Beijing, China aff002;  Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China aff003;  Department of General Practice, Nanjing First Hospital, Nanjing Medical University, Nanjing, China aff004;  Beijing Advanced Innovation Center for Structural Biology, Tsinghua University, Beijing, China aff005
Vyšlo v časopise: Identification of functional cis-acting RNA elements in the hepatitis E virus genome required for viral replication. PLoS Pathog 16(5): e32767. doi:10.1371/journal.ppat.1008488
Kategorie: Research Article
doi: 10.1371/journal.ppat.1008488

Souhrn

There are approximately 20 million events of hepatitis E virus (HEV) infection worldwide annually. The genome of HEV is a single-strand, positive-sense RNA containing 5’ and 3’ untranslated regions and three open reading frames (ORF). HEV genome has 5’ cap and 3’ poly(A) tail to mimic host mRNA to escape the host innate immune surveillance and utilize host translational machineries for viral protein translation. The replication mechanism of HEV is poorly understood, especially how the viral polymerase distinguishes viral RNA from host mRNA to synthesize new viral genomes. We hypothesize that the HEV genome contains cis-acting elements that can be recognized by the virally encoded polymerase as “self” for replication. To identify functional cis-acting elements systematically across the HEV genome, we utilized an ORF1 transcomplementation system. Ultimately, we found two highly conserved cis-acting RNA elements within the ORF1 and ORF2 coding regions that are required for viral genome replication in a diverse panel of HEV genotypes. Synonymous mutations in the cis-acting RNA elements, not altering the ORF1 and ORF2 protein sequences, significantly impaired production of infectious viral particles. Mechanistic studies revealed that the cis-acting elements form secondary structures needed to interact with the HEV ORF1 protein to promote HEV replication. Thus, these cis-acting elements function as a scaffold, providing a specific “signal” that recruits viral and host factors to assemble the viral replication complex. Altogether, this work not only facilitates our understanding of the HEV life cycle and provides novel, RNA-directed targets for potential HEV treatments, but also sheds light on the development of HEV as a therapeutic delivery vector.

Klíčová slova:

Hepatitis E virus – Immunoprecipitation – RNA structure – RNA synthesis – Viral genome – Viral genomics – Viral replication – Viral replication complex


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