Cleavage of oligoribonucleotides by a ribozyme derived from the hepatitis delta virus RNA sequence.
A self-cleaving RNA sequence from hepatitis delta virus was modified to produce a ribozyme capable of catalyzing the cleavage of RNA in an intermolecular (trans) reaction. The delta-derived ribozyme cleaved substrate RNA at a specific site, and the sequence specificity could be altered with mutations in the region of the ribozyme proposed to base pair with the substrate. A substrate target size of approximately 8 nucleotides in length was identified. Octanucleotides containing a single ribonucleotide immediately 5' to the cleavage site were substrates for cleavage, and cleavage activity was significantly reduced only with a guanine base at that position. A deoxyribose 5' to the cleavage site blocked the reaction. These data are consistent with a proposed secondary structure for the self-cleaving form of the hepatitis delta virus ribozyme in which a duplex forms with sequences 3' to the cleavage site, and they support a proposed mechanism in which cleavage involves attack on the phosphorus at the cleavage site by the adjacent 2'-hydroxyl group.
Perrotta
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Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710.
Been
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MD
D001483:Base Sequence; D001665:Binding Sites; D003698:Hepatitis Delta Virus / Q000201:enzymology / Q000235:genetics*; D006571:Heterocyclic Compounds / Q000378:metabolism; D006868:Hydrolysis; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009690:Nucleic Acid Conformation; D009705:Nucleosides / Q000378:metabolism; D009843:Oligoribonucleotides / Q000378:metabolism*; D016337:RNA, Catalytic / Q000378:metabolism*; D012367:RNA, Viral / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013379:Substrate Specificity
D006571:Heterocyclic Compounds; D009705:Nucleosides; D009843:Oligoribonucleotides; D016337:RNA, Catalytic; D012367:RNA, Viral