Journal of Bacteriology, December 1998, p. 6232-6241, Vol. 180, No. 23
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, United Kingdom
Received 14 May 1998/Accepted 10 September 1998
The staphylococcal accessory regulator (encoded by
sarA) is an important global regulator of virulence factor
biosynthesis in Staphylococcus aureus. To further
characterize its role in virulence determinant production, an
sarA knockout mutant was created by insertion of a
kanamycin antibiotic resistance cassette into the sarA
gene. N-terminal sequencing of exoproteins down-regulated by
sarA identified several putative proteases, including a V8 serine protease and a novel metalloprotease, as the major extracellular proteins repressed by sarA. In kinetic studies, the
sarA mutation delays the onset of
-hemolysin (encoded by
hla) expression and reduces levels of hla to
approximately 40% of the parent strain level. Furthermore, SarA plays
a role in signal transduction in response to microaerobic growth since
levels of hla were much lower in a microaerobic environment
than after aerobic growth in the sarA mutant. An exoprotein
exhibiting hemolysin activity on sheep blood, and up-regulated by
sarA independently of the accessory gene regulator (encoded
by agr), was specifically induced microaerobically.
Transcriptional gene fusion and Western analysis revealed that
sarA up-regulates both toxic shock syndrome toxin 1 gene
(tst) expression and staphylococcal enterotoxin B
production, respectively. This study demonstrates the role of
sarA as a signal transduction regulatory component in
response to aeration stimuli and suggests that sarA
functions as a major repressor of protease activity. The possible role
of proteases as regulators of virulence determinant stability is discussed.
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