A NOVEL SUICIDE SUBSTRATE FOR DNA TOPOISOMERASES AND SITE-SPECIFIC RECOMBINASES
A NOVEL SUICIDE SUBSTRATE FOR DNA TOPOISOMERASES AND SITE-SPECIFIC RECOMBINASES
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DOI：
10.1093/nar/23.15.2973
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发表时间：
1995-08-11
影响因子：
14.9
通讯作者：
NASH, HA
中科院分区：
文献类型：
--
作者：
BURGIN, AB;HUIZENGA, BN;NASH, HA
DNA拓扑异构酶和DNA定点重组酶是参与多种细胞过程的重要生物酶。我们发现5‘-桥联硫代磷酸取代了磷酸二酯键，为小牛胸腺拓扑异构酶I和lambda整合酶蛋白(Int)创造了一个有效的自杀底物。虽然桥联硫代磷酸键被这些酶切割，但生成的5‘-巯基不适合后续的连接反应。我们利用Int促进的切割的不可逆性来探索导致lambda整合重组的各个步骤的条件和因素，硫代磷酸底物比传统的自杀底物具有优势，可能是抑制相关细胞酶的有效工具，并为许多其他磷酸转移反应的研究提供了一个独特的工具。
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.