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

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.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Burgin AB Jr,Huizenga BN,Nash HA

doi

10.1093/nar/23.15.2973

subject

Has Abstract

pub_date

1995-08-11 00:00:00

pages

2973-9

issue

15

eissn

0305-1048

issn

1362-4962

pii

5a0177

journal_volume

23

pub_type

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