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Applied and Environmental Microbiology, November 2002, p. 5549-5553, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5549-5553.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Isolation and Molecular Analysis of the Gene Cluster for the Arginine Deiminase System from Streptococcus gordonii DL1

Yiqian Dong, Yi-Ywan M. Chen, Jennifer A. Snyder,{dagger} and R. A. Burne*

Department of Oral Biology, University of Florida, Gainesville, Florida 32610

Received 3 June 2002/ Accepted 14 August 2002

The arginine deiminase (AD) system (ADS) is one of two major ammonia-generating pathways in the oral cavity that play important roles in oral biofilm pH homeostasis and oral biofilm ecology. To initiate a study of the Streptococcus gordonii ADS, the ADS gene cluster was isolated from subgenomic DNA libraries of S. gordonii DL1 by using an arcB-specific probe. Nucleotide sequence analysis revealed six open reading frames (ORFs) that were arranged contiguously; the first five ORFs were transcribed in the same direction, as an apparent operon, and the sixth was transcribed in the opposite direction. The ORFs were found to share significant homologies and to correspond closely in molecular mass to previously characterized arc genes; thus, they were designated arcA (AD), arcB (ornithine carbamyltransferase), arcC (carbamate kinase), arcD (arginine-ornithine antiporter), arcT (dipeptidase), and arcR (regulator). A putative {sigma}70 promoter (ParcA [TTGTGT-N19-TAGAAT]) was mapped 5' to arcA by primer extension, and the expression of ParcA was shown to be inducible by arginine and repressible by glucose, in agreement with AD specific activities measured in the wild-type strain. To investigate the function of ArcR in the differential expression of the arc operon, arcR was insertionally inactivated by a KM resistance marker flanked by T4 transcription/translation termination signals, and the expression of ParcA was monitored by primer extension in the wild-type and ArcR-deficient strains. Lower levels of arcA expression, as well as lower levels of AD activity, were consistently observed in the ArcR-deficient strain compared to wild-type cells, regardless of the growth conditions. Thus, ArcR is a transcriptional activator that is required for induction and optimal expression of the S. gordonii ADS gene cluster.

* Corresponding author. Mailing address: Department of Oral Biology, University of Florida, 1600 SW Archer Rd., P.O. Box 100424, Gainesville, FL 32610. Phone: (352) 392-4370. Fax: (352) 392-7357. E-mail: rburne{at}dental.ufl.edu.

{dagger} Present address: Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201.

Applied and Environmental Microbiology, November 2002, p. 5549-5553, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5549-5553.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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