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Biochemistry. 1992 Jan 14;31(1):16-21.

Cleavage of oligoribonucleotides by a ribozyme derived from the hepatitis delta virus RNA sequence.

Author information

1
Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710.

Abstract

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.

PMID:
1731868
DOI:
10.1021/bi00116a004
[Indexed for MEDLINE]

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