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PMID: 9060427 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Mechanisms of triplex-caused polymerization arrest.

Nucleic acids research ·Vol. 25 ·No. 7 ·1997-04-01 ·Pages 1339-46

Krasilnikov AS, Panyutin IG, Samadashwily GM, Cox R, Lazurkin YS, Mirkin SM

Abstract

Pyrimidine/purine/purine triplexes are known to inhibit DNA polymerization. Here we have studied the mechanisms of this inhibition by comparing the efficiency of Vent DNA polymerase on triplex- and duplex-containing templates at different temperatures, Mg2+concentrations and time intervals with the thermal stability of the corresponding structures. Our results show that triplexes can only be by-passed at temperatures where thermal denaturation initiates, while duplexes, in contrast, are overcome at temperatures where they are quite stable. These results show that DNA polymerase cannot untangle triplex regions within DNA templates and seems to entirely depend on their thermal fluctuations. The high stability of triplexes at physiological temperatures and ambient conditions make them a barrier to polymerization.

MeSH Terms

Base Sequence Biopolymers/metabolism DNA/biosynthesis DNA-Directed DNA Polymerase/metabolism Electrophoresis, Polyacrylamide Gel Gene Targeting Kinetics Magnesium/metabolism Molecular Sequence Data Nucleic Acid Conformation Substrate Specificity Temperature Templates, Genetic

Chemicals

Biopolymers triplex DNA DNA Deep Vent DNA polymerase DNA-Directed DNA Polymerase Magnesium

Authors & Affiliations

6 authors, click to expand affiliations / ORCID

Krasilnikov A S

Department of Genetics, University of Illinois at Chicago, Chicago, IL 60607, USA.

Panyutin I G

Samadashwily G M

Cox R

Lazurkin Y S

Mirkin S M

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Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/25.7.1339

Published

1997-04-01

Pages

1339-46

Language

English

Region

England

NLM ID

0411011

PMCID

PMC146602

Subset

External Links

PubMed source DOI: 10.1093/nar/25.7.1339 PMC Full Text View journal details

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