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J. Biol. Chem., Vol. 276, Issue 12, 9413-9420, March 23, 2001
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From the Laboratory of Molecular Genetics, NIEHS, National
Institutes of Health, Research Triangle Park,
North Carolina 27709
This study examines the interaction between
Escherichia coli MutS,L and E. coli RuvAB
during E. coli RecA-promoted strand exchange. RuvAB is a
branch migration complex that stimulates heterologous strand exchange.
Previous studies indicate that RuvAB increases the rate at which
heteroduplex products are formed by RecA, that RuvA and RuvB are
required for this stimulation, and that RuvAB does not stimulate
homologous strand exchange. This study indicates that MutS,L inhibit
the formation of full-length heteroduplex DNA between M13-fd DNA
in the presence of RuvAB, such that less than 2% of the linear
substrate is converted to product. Inhibition depends on the time at
which MutS,L are added to the reaction and is strongest when MutS,L are
added during initiation. The kinetics of the strand exchange reaction
suggest that MutS,L directly inhibit RuvAB-dependent branch
migration in the absence of RecA. The inhibition requires the formation of base-base mismatches and ATP utilization; no effect on
RuvAB-promoted strand exchange is seen if an ATP-deficient mutant of
MutS (MutS501) is included in the reaction instead of wild-type MutS.
These results are consistent with a role for MutS,L in maintaining
genomic stability and replication fidelity.
Escherichia coli MutS,L Modulate
RuvAB-dependent Branch Migration between Diverged DNA*
and
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Permanent address: Inst. of Biochemistry and Biophysics,
Polish Academy of Science, Pawinskiego 5a, 02-106 Warsaw, Poland.
§
To whom correspondence and reprint requests should be addressed.
Tel.: 919-541-0670; Fax: 919-541-7593; E-mail:
worth@niehs.nih.gov.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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