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Journal of Bacteriology, November 1998, p. 5866-5874, Vol. 180, No. 22
Department of Biochemistry, University of
Wisconsin
Received 18 June 1998/Accepted 11 September 1998
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.
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Escherichia coli DNA Topoisomerase I and Suppression
of Killing by Tn5 Transposase Overproduction:
Topoisomerase I Modulates Tn5 Transposition
Madison, Madison, Wisconsin 53706
*
Corresponding author. Mailing address: Department of
Biochemistry, 433 Babcock Dr., Madison, WI 53706-1544. Phone: (608)
262-3608. Fax: (608) 262-3453. E-mail:
reznikoff{at}biochem.wisc.edu.
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