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A role for the Clp protease in activating Mu-mediated DNA rearrangements.

Department of Biochemistry and Molecular Biology, University of Chicago, Illinois 60637.

ABSTRACT

Bacteriophage Mu, one of the best-characterized mobile genetic elements, can be used effectively to answer fundamental questions about the regulation of biochemical machinery for DNA rearrangement. Previous studies of Mu virulence have implicated the Clp protease in repressor inactivation (V. Geuskens, A. Mhammedi-Alaoui, L. Desmet, and A. Toussaint, EMBO J. 13:5121-5127, 1992). These studies were extended by analyzing the phenotypic consequences of clp alleles in two Escherichia coli systems: (i) the periodic replication of Mudlac transposons in colonies and (ii) the action of a Mu prophage in forming araB-lacZ coding sequence fusions. The clpP::CM mutation, which removes the proteolytic subunit of Clp protease, caused a drastic reduction in Mu activity in both systems. The clpA::Tn10 mutation, which removes a regulatory subunit of Clp protease, altered the timing of Mu activity in both systems. A clpA deletion reduced the extent of Mudlac replication in colonies. These results point to temporal changes in Clp proteolysis of the Mucts62 repressor as a key molecular event in the regulation of one class of genomic change in E. coli.

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