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Journal of Bacteriology, November 2000, p. 6014-6026, Vol. 182, No. 21
Department of Microbiology, College of
Physicians and Surgeons, Columbia University, New York, New York
10032
Received 10 April 2000/Accepted 5 August 2000
Replication of the broad-host-range, IncP
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Incompatibility Protein IncC and Global Regulator KorB Interact
in Active Partition of Promiscuous Plasmid RK2

and
plasmid RK2 requires
two plasmid loci: trfA, the replication initiator gene, and oriV, the origin of replication. While these determinants
are sufficient for replication in a wide variety of bacteria, they do
not confer the stable maintenance of parental RK2 observed in its
hosts. The product of the incC gene has been proposed to function in the stable maintenance of RK2 because of its relatedness to
the ParA family of ATPases, some of which are known to be involved in
the active partition of plasmid and chromosomal DNA. Here we show that
IncC has the properties expected of a component of an active partition
system. The smaller polypeptide product of incC (IncC2)
exhibits a strong, replicon-independent incompatibility phenotype with
RK2. This incompatibility phenotype requires the global transcriptional
repressor, KorB, and the target for incC-mediated incompatibility is a KorB-binding site (OB). We found that
KorB and IncC interact in vivo by using the yeast two-hybrid system and
in vitro by using partially purified proteins. Elevated expression of
the incC and korB genes individually has no
obvious effect on Escherichia coli cell growth, but their
simultaneous overexpression is toxic, indicating a possible interaction
of IncC-KorB complexes with a vital host target. A region of RK2
bearing incC, korB, and multiple KorB-binding
sites is able to stabilize an unstable, heterologous plasmid in an
incC-dependent manner. Finally, elevated levels of IncC2
cause RK2 to aggregate, indicating a possible role for IncC in plasmid
pairing. These findings demonstrate that IncC, KorB, and at least one
KorB-binding site are components of an active partition system for the
promiscuous plasmid RK2.
*
Corresponding author. Mailing address: Department of
Microbiology, College of Physicians and Surgeons, Columbia University, 701 W. 168th St., New York, NY 10032. Phone: (212) 305-3425. Fax: (212)
305-1468. E-mail: figurski{at}cuccfa.ccc.columbia.edu.
Present address: Department of Microbiology, University of Texas
Southwestern Medical Center, Dallas, TX 75390.
Present address: Department of Microbiology and Immunology, Albert
Einstein College of Medicine, Bronx, NY 10461.
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