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PMID: 2467291 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Site-selective cleavage of structured RNA by a staphylococcal nuclease-DNA hybrid.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 6 ·1989-03-00 ·Pages 1766-70

Zuckermann RN, Schultz PG

Abstract

A hybrid enzyme consisting of an oligodeoxyribonucleotide fused to a unique site on staphylococcal nuclease site-selectively cleaves a number of natural RNAs including Escherichia coli M1 RNA (377 bases), 16S rRNA (1542 bases), and yeast tRNA(Phe). The oligonucleotide directs the nuclease activity of the enzyme to the nucleotides directly adjacent to the complementary target sequence on the substrate RNA. In the case of M1 RNA, hydrolysis occurs primarily at one phosphodiester bond, converting 50% of the starting material to product. Furthermore, the reaction products can be enzymatically manipulated: tRNA(Phe) was cleaved in the anticodon region and was religated to form the full-length tRNA in high yield. Because the specificity of these hybrid enzymes can be easily altered, they should prove to be useful tools for probing RNA structure and function.

MeSH Terms

Base Sequence Binding Sites DNA/metabolism Drug Stability Escherichia coli Hydrolysis Micrococcal Nuclease/metabolism Molecular Sequence Data Nucleic Acid Hybridization RNA/metabolism RNA, Bacterial/metabolism RNA, Fungal/metabolism RNA, Ribosomal, 16S/metabolism RNA, Transfer, Phe/metabolism Recombinant Fusion Proteins/metabolism Recombinant Proteins/metabolism

Chemicals

RNA, Bacterial RNA, Fungal RNA, Ribosomal, 16S RNA, Transfer, Phe Recombinant Fusion Proteins Recombinant Proteins RNA DNA Micrococcal Nuclease

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Zuckermann R N

Department of Chemistry, University of California, Berkeley 94720.

Schultz P G

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Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.86.6.1766

Published

1989-03-00

Pages

1766-70

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC286785

Subset

External Links

PubMed source DOI: 10.1073/pnas.86.6.1766 PMC Full Text View journal details

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Site-selective cleavage of structured RNA by a staphylococcal nuclease-DNA hybrid.

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