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PMID: 2308821 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Site-specific inhibition of EcoRI restriction/modification enzymes by a DNA triple helix.

Nucleic acids research ·Vol. 18 ·No. 1 ·1990-01-11 ·Pages 157-61

Hanvey JC, Shimizu M, Wells RD

Abstract

The ability of oligopyrimidines to inhibit, through triple helix formation, the specific protein-DNA interactions of the EcoRI restriction and modification enzymes (EcoRI and MEcoRI) with their recognition sequence (GAATTC) was studied. The oligonucleotides (CTT)4 and (CTT)8 formed triplexes in plasmids at (GAA)n repeats containing EcoRI sites. Cleavage and methylation of EcoRI sites within these sequences were specifically inhibited by the oligonucleotides, whereas an EcoRI site adjacent to a (GAA)n sequence was inhibited much less. Also, other EcoRI sites within the plasmid, or in exogenously added lambda DNA, were not inhibited. These results demonstrate the potential of using triplex-forming oligonucleotides to block protein-DNA interactions at specific sites, and thus this technique may be useful in chromosome mapping and in the modulation of gene expression.

MeSH Terms
DNA Modification Methylases/antagonists & inhibitors Deoxyribonuclease EcoRI/antagonists & inhibitors Methylation Nucleic Acid Conformation Oligodeoxyribonucleotides/pharmacology Plasmids Sulfuric Acid Esters
Chemicals
Oligodeoxyribonucleotides Sulfuric Acid Esters DNA Modification Methylases Deoxyribonuclease EcoRI dimethyl sulfate
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hanvey J C
Department of Biochemistry, School of Medicine, University of Alabama, Birmingham 35294.
Shimizu M
Wells R D
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/18.1.157
Published
1990-01-11
Pages
157-61
Language
English
Region
England
NLM ID
0411011
PMCID
PMC330216
Subset
IM
Grants
NCI NIH HHS · CA13148 · United States
NIGMS NIH HHS · GM30822 · United States
External Links
PubMed source DOI: 10.1093/nar/18.1.157 PMC Full Text View journal details
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