Affiliation:
1. Department of Biochemistry, University of Wisconsin—Madison, Madison, Wisconsin 53706
Abstract
ABSTRACT
Tn
5
transposase (Tnp) overproduction is lethal to
Escherichia coli
. Genetic evidence suggested that this killing involves titration of
E. coli
topoisomerase I (Topo I). Here, we present biochemical evidence that supports this model. Tn
5
Tnp copurifies with Topo I while nonkilling derivatives of Tnp, Δ37Tnp and Δ55Tnp (Inhibitor [Inh]), show reduced affinity or no affinity, respectively, for Topo I. In agreement with these results, the presence of Tnp, but not Δ37 or Inh derivatives of Tnp, inhibits the DNA relaxation activity of Topo I in vivo as well as in vitro. Other proteins, including RNA polymerase, are also found to copurify with Tnp. For RNA polymerase, reduced copurification with Tnp is observed in extracts from a
topA
mutant strain, suggesting that RNA polymerase interacts with Topo I and not Tnp.
Publisher
American Society for Microbiology
Subject
Molecular Biology,Microbiology
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