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Journal of Bacteriology, May 1999, p. 2863-2871, Vol. 181, No. 9
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Regulation of Expression of the Fructan Hydrolase
Gene of Streptococcus mutans GS-5 by Induction and
Carbon Catabolite Repression
Robert A.
Burne,1,2,*
Zezhang
Thomas
Wen,1
Yi-Ywan M.
Chen,1,2 and
Jana E. C.
Penders1
Center for Oral
Biology1 and Department of Microbiology
and Immunology,2 University of Rochester
School of Medicine and Dentistry, Rochester, New York 14642
Received 29 September 1998/Accepted 25 February 1999
The polymers of fructose, levan and inulin, as well as sucrose and
raffinose, are substrates for the product of the fruA gene of Streptococcus mutans GS-5. The purpose of this study was
to characterize the DNA immediately flanking fruA, to
explore the regulation of expression of fruA by the
carbohydrate source, and to begin to elucidate the molecular basis for
differential expression of the gene. Located 3' to fruA was
an open reading frame (ORF) with similarity to
-fructosidases which
was cotranscribed with fruA. A transcriptional initiation
site, located an appropriate distance from an extended
10-like
promoter, was mapped at 165 bp 5' to the fruA structural
gene. By the use of computer algorithms, two overlapping, stable
stem-loop sequences with the potential to function as rho-independent
terminators were found in the 5' untranslated region. Catabolite
response elements (CREs), which have been shown to govern carbon
catabolite repression (CCR) by functioning as negative cis
elements in gram-positive bacteria, were located close to the promoter.
The levels of production of fruA mRNA and FruA were
elevated in cells growing on levan, inulin, or sucrose as the sole
carbohydrate source, and repression was observed when cells were grown
on readily metabolizable hexoses. Deletion derivatives containing
fusions of fruA promoter regions, lacking sequences 5' or
3' to the promoter, and a promoterless chloramphenicol
acetyltransferase gene were used (i) to demonstrate the functionality
of the promoter mapped by primer extension, (ii) to demonstrate that
CCR of the fru operon requires the CRE that is located 3'
to the promoter region, and (iii) to provide preliminary evidence that
supports the involvement of an antitermination mechanism in
fruA induction.
*
Corresponding author. Mailing address: Center for Oral
Biology, University of Rochester School of Medicine and Dentistry, 601 Elmwood Ave., Rochester, NY 14642. Phone: (716) 275-0381. Fax: (716)
473-2679. E-mail: robert_burne{at}urmc.rochester.edu.
Journal of Bacteriology, May 1999, p. 2863-2871, Vol. 181, No. 9
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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