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Cell ; 187(11): 2735-2745.e12, 2024 May 23.
Artículo en Inglés | MEDLINE | ID: mdl-38723628

RESUMEN

Hepatitis B virus (HBV) is a small double-stranded DNA virus that chronically infects 296 million people. Over half of its compact genome encodes proteins in two overlapping reading frames, and during evolution, multiple selective pressures can act on shared nucleotides. This study combines an RNA-based HBV cell culture system with deep mutational scanning (DMS) to uncouple cis- and trans-acting sequence requirements in the HBV genome. The results support a leaky ribosome scanning model for polymerase translation, provide a fitness map of the HBV polymerase at single-nucleotide resolution, and identify conserved prolines adjacent to the HBV polymerase termination codon that stall ribosomes. Further experiments indicated that stalled ribosomes tether the nascent polymerase to its template RNA, ensuring cis-preferential RNA packaging and reverse transcription of the HBV genome.


Asunto(s)
Virus de la Hepatitis B , Transcripción Reversa , Humanos , Genoma Viral/genética , Virus de la Hepatitis B/genética , Mutación , Ribosomas/metabolismo , ARN Viral/genética , ARN Viral/metabolismo , Línea Celular
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