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Journal of Bacteriology, April 2000, p. 1794-1801, Vol. 182, No. 7
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
The Staphylococcus aureus lrgAB Operon
Modulates Murein Hydrolase Activity and Penicillin Tolerance
Kajetan H.
Groicher,
Brian A.
Firek,
David F.
Fujimoto, and
Kenneth W.
Bayles*
Department of Microbiology, Molecular Biology
and Biochemistry, University of Idaho, Moscow, Idaho 83844-3052
Received 23 September 1999/Accepted 7 January 2000
Previous studies in our laboratory have shown that the
Staphylococcus aureus LytSR two-component regulatory system
affects murein hydrolase activity and autolysis. A LytSR-regulated
dicistronic operon has also been identified and shown to encode two
potential membrane-associated proteins, designated LrgA and LrgB,
hypothesized to be involved in the control of murein hydrolase
activity. In the present study, a lrgAB mutant strain was
generated and analyzed to test this hypothesis. Zymographic and
quantitative analysis of murein hydrolase activity revealed that the
lrgAB mutant produced increased extracellular murein
hydrolase activity compared to that of the wild-type strain.
Complementation of the lrgAB defect by providing the
lrgAB genes in trans restored the wild-type
phenotype, indicating that these genes confer negative control on
extracellular murein hydrolase activity. In addition to these effects,
the influence of the lrgAB mutation on penicillin-induced
lysis and killing was examined. These studies demonstrated that the
lrgAB mutation enhanced penicillin-induced killing of cells
approaching the stationary phase of growth, the time at which the
lrgAB operon was shown to be maximally expressed. This
effect of the lrgAB mutation on penicillin-induced killing
was shown to be independent of cell lysis. In contrast, the
lrgAB mutation did not affect penicillin-induced killing of
cells growing in early-exponential phase, a time in which
lrgAB expression was shown to be minimal. However,
expression of the lrgAB operon in early-exponential-phase
cells inhibited penicillin-induced killing, again independent of cell
lysis. The data generated by this study suggest that penicillin-induced
killing of S. aureus involves a novel regulator of murein
hydrolase activity.
*
Corresponding author. Mailing address: Department of
Microbiology, Molecular Biology and Biochemistry, College of
Agriculture, University of Idaho, Moscow, ID 83844-3052. Phone: (208)
885-7164. Fax: (208) 885-6518. E-mail: kbayles{at}uidaho.edu.
Journal of Bacteriology, April 2000, p. 1794-1801, Vol. 182, No. 7
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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