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PMID: 7659520 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A novel suicide substrate for DNA topoisomerases and site-specific recombinases.

Nucleic acids research ·Vol. 23 ·No. 15 ·1995-08-11 ·Pages 2973-9

Burgin AB, Huizenga BN, Nash HA

Abstract

DNA topoisomerases and DNA site-specific recombinases are biologically important enzymes involved in a diverse set of cellular processes. We show that replacement of a phosphodiester linkage by a 5'-bridging phosphorothioate linkage creates an efficient suicide substrate for calf thymus topoisomerase I and lambda integrase protein (Int). Although the bridging phosphorothioate linkage is cleaved by these enzymes, the 5'-sulfhydryl which is generated is not competent for subsequent ligation reactions. We use the irreversibility of Int-promoted cleavage to explore conditions and factors that contribute to various steps of lambda integrative recombination. The phosphorothioate substrates offer advantages over conventional suicide substrates, may be potent tools for inhibition of the relevant cellular enzymes and represent a unique tool for the study of many other phosphoryl transfer reactions.

MeSH Terms
Animals Bacterial Proteins/metabolism Base Sequence Cattle DNA/chemistry,metabolism DNA Nucleotidyltransferases/antagonists & inhibitors,metabolism DNA Topoisomerases, Type I/metabolism DNA, Viral/metabolism DNA-Binding Proteins/metabolism Integrases Integration Host Factors Molecular Sequence Data Polydeoxyribonucleotides/chemical synthesis,chemistry,metabolism Substrate Specificity Thionucleotides/metabolism Topoisomerase I Inhibitors Virus Integration/physiology
Chemicals
Bacterial Proteins DNA, Viral DNA-Binding Proteins Integration Host Factors Polydeoxyribonucleotides Thionucleotides Topoisomerase I Inhibitors DNA DNA Nucleotidyltransferases Integrases DNA Topoisomerases, Type I
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Burgin A B
Laboratory of Molecular Biology, National Institute of Mental Health, Bethesda, MD 20892, USA.
Huizenga B N
Nash H A
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/23.15.2973
Published
1995-08-11
Pages
2973-9
Language
English
Region
England
NLM ID
0411011
PMCID
PMC307138
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/23.15.2973 PMC Full Text View journal details
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