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PMID: 2555787 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

RNase H cleavage of RNA hybridized to oligonucleotides containing methylphosphonate, phosphorothioate and phosphodiester bonds.

Nucleic acids research ·Vol. 17 ·No. 22 ·1989-11-25 ·Pages 9193-204

Furdon PJ, Dominski Z, Kole R

Abstract

Three types of 14-mer oligonucleotides were hybridized to human beta-globin pre-mRNA and the resultant duplexes were tested for susceptibility to cleavage by RNase H from E. coli or from HeLa cell nuclear extract. The oligonucleotides contained normal deoxynucleotides, phosphorothioate analogs alternating with normal deoxynucleotides, or one to six methylphosphonate deoxynucleosides. Duplexes formed with deoxyoligonucleotides or phosphorothioate analogs were susceptible to cleavage by RNase H from both sources, whereas a duplex formed with an oligonucleotide containing six methylphosphonate deoxynucleosides alternating with normal deoxynucleotides was resistant. Susceptibility to cleavage by RNase H increased parallel to a reduction in the number of methylphosphonate residues in the oligonucleotide. Stability of the oligonucleotides in the nuclear extract from HeLa cells was also tested. Whereas deoxyoligonucleotides were rapidly degraded, oligonucleotides containing alternating methylphosphonate residues remained unchanged after 70 minutes of incubation. Other oligonucleotides exhibited intermediate stability.

MeSH Terms
Base Sequence Cell Nucleus/enzymology Endoribonucleases/metabolism Escherichia coli/enzymology Globins/genetics HeLa Cells/enzymology Humans Kinetics Molecular Sequence Data Nucleic Acid Hybridization Oligodeoxyribonucleotides/chemical synthesis Organophosphorus Compounds RNA Precursors/genetics Ribonuclease H Substrate Specificity Thionucleotides
Chemicals
Oligodeoxyribonucleotides Organophosphorus Compounds RNA Precursors Thionucleotides methylphosphonic acid Globins Endoribonucleases Ribonuclease H
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Furdon P J
Department of Pharmacology, University of North Carolina, Chapel Hill 27599.
Dominski Z
Kole R
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/17.22.9193
Published
1989-11-25
Pages
9193-204
Language
English
Region
England
NLM ID
0411011
PMCID
PMC335124
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/17.22.9193 PMC Full Text View journal details
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