Escherichia coli DNA topoisomerase I and suppression of killing by Tn5 transposase overproduction: topoisomerase I modulates Tn5 transposition.
Tn5 transposase (Tnp) overproduction is lethal to Escherichia coli. The overproduction causes cell filamentation and abnormal chromosome segregation. Here we present three lines of evidence strongly suggesting that Tnp overproduction killing is due to titration of topoisomerase I. First, a suppressor mutation of transposase overproduction killing, stkD10, is localized in topA (the gene for topoisomerase I). The stkD10 mutant has the following characteristics: first, it has an increased abundance of topoisomerase I protein, the topoisomerase I is defective for the DNA relaxation activity, and DNA gyrase activity is reduced; second, the suppressor phenotype of a second mutation localized in rpoH, stkA14 (H. Yigit and W. S. Reznikoff, J. Bacteriol. 179:1704-1713, 1997), can be explained by an increase in topA expression; and third, overexpression of wild-type topA partially suppresses the killing. Finally, topoisomerase I was found to enhance Tn5 transposition up to 30-fold in vivo.
Yigit
H
H
Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
Reznikoff
W S
WS
D002874:Chromosome Mapping; D004264:DNA Topoisomerases, Type I / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D004250:DNA Topoisomerases, Type II / Q000096:biosynthesis / Q000378:metabolism; D004269:DNA, Bacterial / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D010641:Phenotype; D012333:RNA, Messenger; D019895:Transposases
D004269:DNA, Bacterial; D012333:RNA, Messenger; D019895:Transposases; D004264:DNA Topoisomerases, Type I; D004250:DNA Topoisomerases, Type II