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 Resjournal_title
Nucleic acids researchauthors
Burgin AB Jr,Huizenga BN,Nash HAdoi
10.1093/nar/23.15.2973subject
Has Abstractpub_date
1995-08-11 00:00:00pages
2973-9issue
15eissn
0305-1048issn
1362-4962pii
5a0177journal_volume
23pub_type
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