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J Bacteriol, May 1998, p. 2623-2629, Vol. 180, No. 10
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
The L-Isoaspartyl Protein Repair
Methyltransferase Enhances Survival of Aging Escherichia
coli Subjected to Secondary Environmental Stresses
Jonathan E.
Visick,
Hui
Cai, and
Steven
Clarke*
Department of Chemistry and Biochemistry and
the Molecular Biology Institute, University of California, Los
Angeles, California 90095-1569
Received 28 January 1998/Accepted 11 March 1998
Like its homologs throughout the biological world, the
L-isoaspartyl protein repair methyltransferase of
Escherichia coli, encoded by the pcm gene, can
convert abnormal L-isoaspartyl residues in proteins (which
form spontaneously from asparaginyl or aspartyl residues) to normal
aspartyl residues. Mutations in pcm were reported to
greatly reduce survival in stationary phase and when cells were
subjected to heat or osmotic stresses (C. Li and S. Clarke, Proc. Natl.
Acad. Sci. USA 89:9885-9889, 1992). However, we subsequently demonstrated that those strains had a secondary mutation in
rpoS, which encodes a stationary-phase-specific sigma
factor (J. E. Visick and S. Clarke, J. Bacteriol. 179:4158-4163,
1997). We now show that the rpoS mutation, resulting in a
90% decrease in HPII catalase activity, can account for the previously
observed phenotypes. We further demonstrate that a new pcm
mutant lacks these phenotypes. Interestingly, the newly constructed
pcm mutant, when maintained in stationary phase for
extended periods, is susceptible to environmental stresses, including
exposure to methanol, oxygen radical generation by paraquat, high salt
concentrations, and repeated heating to 42°C. The pcm
mutation also results in a competitive disadvantage in stationary-phase
cells. All of these phenotypes can be complemented by a functional
pcm gene integrated elsewhere in the chromosome. These data
suggest that protein denaturation and isoaspartyl formation may act
synergistically to the detriment of aging E. coli and that
the repair methyltransferase can play a role in limiting the
accumulation of the potentially disruptive isoaspartyl residues in
vivo.
*
Corresponding author. Mailing address: Dept. of
Chemistry and Biochemistry, Box 951569, Los Angeles, CA 90095-1569. Phone: (310) 825-8754. Fax: (310) 825-1968. E-mail:
clarke{at}ewald.mbi.ucla.edu.
J Bacteriol, May 1998, p. 2623-2629, Vol. 180, No. 10
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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