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PMID: 8262040 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Suicidal nucleotide sequences for DNA polymerization.

The EMBO journal ·Vol. 12 ·No. 13 ·1993-12-15 ·Pages 4975-83

Samadashwily GM, Dayn A, Mirkin SM

Abstract

Studying the activity of T7 DNA polymerase (Sequenase) on open circular DNAs, we observed virtually complete termination within potential triplex-forming sequences. Mutations destroying the triplex potential of the sequences prevented termination, while compensatory mutations restoring triplex potential restored it. We hypothesize that strand displacement during DNA polymerization of double-helical templates brings three DNA strands (duplex DNA downstream of the polymerase plus a displaced overhang) into close proximity, provoking triplex formation, which in turn prevents further DNA synthesis. Supporting this idea, we found that Sequenase is unable to propagate through short triple-helical stretches within single-stranded DNA templates. Thus, DNA polymerase, by inducing triplex formation at specific sequences in front of the replication fork, causes self-termination. Possible biological implications of such 'conformational suicide' are discussed. Our data also provide a novel way to target DNA polymerases at specific sequences using triplex-forming oligonucleotides.

MeSH Terms
Base Sequence DNA/biosynthesis DNA, Single-Stranded/metabolism DNA-Directed DNA Polymerase/metabolism Molecular Sequence Data Nucleic Acid Conformation Structure-Activity Relationship Substrate Specificity Templates, Genetic
Chemicals
DNA, Single-Stranded DNA bacteriophage T7 induced DNA polymerase DNA-Directed DNA Polymerase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Samadashwily G M
Department of Genetics, University of Illinois at Chicago 60612.
Dayn A
Mirkin S M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1993-12-15
Pages
4975-83
Language
English
Region
England
NLM ID
8208664
PMCID
PMC413758
Subset
IM
Grants
NIGMS NIH HHS · R55GM46405-01A1 · United States
External Links
PubMed source PMC Full Text View journal details
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