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Journal of Bacteriology, June 2001, p. 3372-3382, Vol. 183, No. 11
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.11.3372-3382.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Characterization of fsr, a Regulator
Controlling Expression of Gelatinase and Serine Protease in
Enterococcus faecalis OG1RF
Xiang
Qin,1,2
Kavindra V.
Singh,1,2
George M.
Weinstock,1,2 and
Barbara E.
Murray1,2,3,*
Division of Infectious Diseases, Department
of Medicine,1 Department of Microbiology
and Molecular Genetics,3 and Center for
the Study of Emerging and Re-emerging
Pathogens,2 University of Texas Medical
School, Houston, Texas 77030
Received 8 November 2000/Accepted 13 March 2001
We have previously identified a locus, fsr, a
homologue of staphylococcal agr loci, which positively
regulates the expression of gelatinase and serine protease (encoded by
gelE and sprE, respectively) in
Enterococcus faecalis OG1RF. The expression of the three
genes in the fsr locus, fsrA,
fsrB, and fsrC, appears to be
autoregulated, and we have shown that mutants with insertion
disruptions in each of these three genes were significantly attenuated
in a mouse peritonitis model compared to the parent strain. In the
present study, we showed that fsrB and
fsrC are highly expressed in the postexponential growth
phase and that their expression is cell density dependent. Reverse
transcriptase PCR using primers covering the intergenic regions
in the fsr/gelE loci confirmed that
fsrB and fsrC, as well as
gelE and sprE, are cotranscribed. We also showed, using a nonpolar fsrB deletion mutant, that
fsrB, the homologue of agrB of
staphylococci with unknown function, is required for the regulatory
function of fsr. Primer extension and analysis of
transcriptional fusions indicated the presence of promoters immediately
upstream of fsrA, of fsrB, and of
gelE and that the fsrB and
gelE promoters are fsr dependent, while
the fsrA promoter is an fsr-independent
weak constitutive promoter. Two conserved 7-bp direct repeats were
found immediately upstream of the fsrB and
gelE promoters, similar to the repeats found upstream of
P2 and P3 promoters of the agr locus; deletions and
mutations in the repeated sequences completely abolished the
fsrB and gelE promoter activities,
suggesting that the repeats are important for the regulatory function
in the fsrB and gelE promoter regions.
*
Corresponding author. Mailing address: Center for the
Study of Emerging and Re-emerging Pathogens, Division of Infectious Diseases, Department of Medicine, University of Texas Medical School at
Houston, 6431 Fannin St., Houston, TX 77030. Phone: (713) 500-6767. Fax: (713) 500-5495. E-mail:
Barbara.E.Murray{at}uth.tmc.edu.
Journal of Bacteriology, June 2001, p. 3372-3382, Vol. 183, No. 11
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.11.3372-3382.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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