Alteration of DNA primary structure by DNA topoisomerase I. Isolation of the covalent topoisomerase I-DNA binary complex in enzymatically competent form

KA Henningfeld, T Arslan, SM Hecht

Index: Henningfeld, Kristine A.; Arslan, Tuncer; Hecht, Sidney M. Journal of the American Chemical Society, 1996 , vol. 118, # 47 p. 11701 - 11714

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Citation Number: 36

Abstract

DNA ligation by DNA topoisomerase I was investigated employing synthetic DNA substrates containing a single strand nick. Site-specific cleavage of the DNA by topoisomerase I in proximity to the nick resulted in uncoupling of the cleavage and ligation reactions of the enzyme, thereby trapping the covalent enzyme-DNA intermediate. DNA cleavage could be reversed by the addition of acceptor oligonucleotides containing a free 5'-OH group and ...