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J. Biol. Chem., Vol. 276, Issue 33, 31053-31058, August 17, 2001
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From the Previous biochemical analysis of
Escherichia coli methyl-directed mismatch repair implicates
three redundant single-strand DNA-specific exonucleases (RecJ, ExoI,
and ExoVII) and at least one additional unknown exonuclease in the
excision reaction (Cooper, D. L., Lahue, R. S., and Modrich,
P. (1993) J. Biol. Chem. 268, 11823-11829). We show
here that ExoX also participates in methyl-directed mismatch repair.
Analysis of the reaction with crude extracts and purified components
demonstrated that ExoX can mediate repair directed from a strand signal
3' of a mismatch. Whereas extracts of all possible single, double, and
triple exonuclease mutants displayed significant residual mismatch
repair, extracts deficient in RecJ, ExoI, ExoVII, and ExoX exonucleases
were devoid of normal repair activity. The RecJ
Redundant Exonuclease Involvement in Escherichia
coli Methyl-directed Mismatch Repair*
§,
,,**, and
Department of Biology and
Rosenstiel Basic Medical Sciences Research Center, Brandeis
University, Waltham, Massachusetts 02254-9110 and the
¶ Department of Biochemistry and Howard Hughes Medical
Institute, Duke University Medical Center, Durham, North Carolina
27710
ExoVII ExoI ExoX strain
displayed a 7-fold increase in mutation rate, a significant increase,
but less than that observed for other blocks of the mismatch repair
pathway. This elevation is epistatic to deficiency for MutS, suggesting
an effect via the mismatch repair pathway. Our other work (Burdett, V.,
Baitinger, C., Viswanathan, M., Lovett, S. T., and Modrich,
P. (2001) Proc. Natl. Acad. Sci. U. S. A. 98, 6765-6770) suggests that mutants are under-recovered in the exonuclease-deficient strain due to loss of viability that is triggered
by mismatched base pairs in this genetic background. The availability
of any one exonuclease is enough to support full mismatch correction,
as evident from the normal mutation rates of all triple mutants.
Because three of these exonucleases possess a strict polarity of
digestion, this suggests that mismatch repair can occur exclusively
from a 3' or a 5' direction to the mismatch, if necessary.
*
This work was supported by Grants GM43889 (to S. T. L.),
GM07122 (to M. V.), and GM23719 (to P. M.) from the General Medical Institute of the National Institutes of Health.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.
To whom correspondence should be addressed: Rosenstiel Basic
Medical Sciences Research Center, MS029, Waltham, MA 02454-9110. Tel.:
781-736-2497; Fax: 781-736-2405; E-mail lovett@brandeis.edu.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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