title: Inhibition of Mycobacterium smegmatis topoisomerase I by specific oligonucleotides.
abstract: DNA topoisomerase I from Mycobacterium smegmatis unlike many other type I topoisomerases is a site specific DNA binding protein. We have investigated the sequence specific DNA binding characteristics of the enzyme using specific oligonucleotides of varied length. DNA binding, oligonucleotide competition and covalent complex assays show that the substrate length requirement for interaction is much longer ( approximately 20 nucleotides) in contrast to short length substrates (eight nucleotides) reported for Escherichia coli topoisomerase I and III. P1 nuclease and KMnO(4) footprinting experiments indicate a large protected region spanning about 20 nucleotides upstream and 2-3 nucleotides downstream of the cleavage site. Binding characteristics indicate that the enzyme interacts efficiently with both single-stranded and double-stranded substrates containing strong topoisomerase I sites (STS), a unique property not shared by any other type I topoisomerase. The oligonucleotides containing STS effectively inhibit the M. smegmatis topoisomerase I DNA relaxation activity.
authors: Microbiology and Cell Biology Department, Indian Institute of Science, 560012, Bangalore, India.
authors: T
authors: T
authors: Bhaduri
authors: S
authors: S
authors: Basak
authors: D
authors: D
authors: Sikder
authors: V
authors: V
authors: Nagaraja
mesh_terms: D001667:Binding, Competitive; D004247:DNA / Q000378:metabolism*; D018983:DNA Footprinting; D004264:DNA Topoisomerases, Type I / Q000378:metabolism; D020102:Mycobacterium smegmatis / Q000201:enzymology*; D009841:Oligonucleotides / Q000378:metabolism / Q000494:pharmacology*; D013379:Substrate Specificity; D059004:Topoisomerase I Inhibitors
chemical_list: D009841:Oligonucleotides; D059004:Topoisomerase I Inhibitors; D004247:DNA; D004264:DNA Topoisomerases, Type I