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Journal of Bacteriology, October 2001, p. 5964-5973, Vol. 183, No. 20
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.20.5964-5973.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

uvrA Is an Acid-Inducible Gene Involved in the Adaptive Response to Low pH in Streptococcus mutans

Michael N. Hanna,1 Ronald J. Ferguson,2 Yung-Hua Li,1 and Dennis G. Cvitkovitch1,*

Dental Research Institute, University of Toronto, Toronto, Ontario, Canada M5G 1G6,1 and Shands Cancer Center, University of Florida, Gainesville, Florida 32610-02322

Received 30 April 2001/Accepted 24 July 2001

The pH-inducible acid tolerance response (ATR) is believed to play a major role in acid adaptation and virulence of Streptococcus mutans. To study this phenomenon in S. mutans JH1005, differential display PCR was used to identify and clone 13 cDNA products that had increased expression in response to pH 5.0 compared to that of pH 7.5-grown cells. One of these products, confirmed to be pH inducible by RNA dot blot and reverse transcription-PCR analyses, had 67% identity to a uvrA-UV repair excinuclease gene in Bacillus subtilis. Further sequence analysis of the uvrA homologue using the S. mutans genome database revealed that the complete gene was encoded in an open reading frame (ORF) of 2,829 bp (944 amino acids; 104.67 kDa). Immediately 3' of uvrA was an ORF encoding a putative aminopeptidase gene (pepP). uvrA knockouts were constructed in S. mutans strains JH1005, NG8, and UA159 using allelic-exchange mutagenesis, replacing the entire gene with an erythromycin resistance cassette. As with uvrA mutants in other bacteria, the S. mutans uvrA mutants were extremely sensitive to UV irradiation. The uvrA mutant of S. mutans JH1005 was also more sensitive than the wild type to growth at pH 5.0, showing a 15% reduction in growth rate and a 14% reduction in final resting culture density. Acid-adapted S. mutans JH1005 uvrA mutants were shown to be more resistant to UV irradiation than was the parent but were unable to survive exposure to a killing pH of 3.0. Moreover, agarose gel electrophoretic analysis of chromosomal DNA isolated from uvrA-deficient cells exposed to low pH demonstrated more DNA damage than that for the wild-type strain. Here we suggest that uvrA and the nucleotide excision repair pathway are involved in the repair of acid-induced DNA damage and are associated with successful adaptation of S. mutans to low pH.

* Corresponding author. Mailing address: Rm. 449A, Dental Research Institute, University of Toronto, 124 Edward St., Toronto, Ontario, Canada M5G 1G6. Phone: (416) 979-4917, ext. 4592. Fax: (416) 979-4936. E-mail: dennis.cvitkovitch{at}utoronto.ca.

Journal of Bacteriology, October 2001, p. 5964-5973, Vol. 183, No. 20
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.20.5964-5973.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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