A pseudoknot-like structure required for efficient self-cleavage of hepatitis delta virus RNA.
Hepatitis delta virus genomic and antigenomic RNAs contain a self-cleavage site hypothesized to function in processing the viral RNA during replication. Self-cleavage requires only a divalent cation and is mediated at the genomic site by a sequence of less than 85 nucleotides. We propose that the genomic self-cleaving sequence element and a corresponding sequence from the anti-genomic RNA could generate related secondary structures. The region of the antigenomic sequence, predicted from the proposed structure, was synthesized and shown to be sufficient for self-cleavage. Evidence for two stems which form a tertiary interaction was obtained by site-specific mutagenesis of the antigenomic sequence. Efficient self-cleavage in 10 M formamide or 5 M urea, also a property of the genomic sequence, was dependent on base-pairing in both stems. But in the absence of denaturants, the stem distal to the site of cleavage was not required, suggesting that the tertiary interaction stabilizes the structure required for self-cleavage.
Perrotta
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AT
Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710.
Been
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MD
D001483:Base Sequence; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D003698:Hepatitis Delta Virus / Q000235:genetics / Q000378:metabolism*; D006860:Hydrogen Bonding; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D015394:Molecular Structure; D016337:RNA, Catalytic / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism* / Q000648:ultrastructure; D013329:Structure-Activity Relationship; D014779:Virus Replication
D016337:RNA, Catalytic; D012367:RNA, Viral