Characterization of cis-acting sites controlling arginine deiminase gene expression in Streptococcus gordonii.
The arginine deiminase system (ADS) is responsible for the production of ornithine, CO2, ammonia, and ATP from arginine. The ADS of the oral bacterium Streptococcus gordonii plays major roles in physiologic homeostasis, acid tolerance, and oral biofilm ecology. To further our understanding of the transcriptional regulation of the ADS (arc) operon, the binding of the ArcR transcriptional activator, which governs expression of the ADS in response to arginine, was investigated by DNase I protection and gel mobility shift assays. An ArcR binding sequence was found that was 27 bp in length and had little sequence similarity to binding sites of other arginine metabolism regulators. The presence of arginine at physiologically relevant concentrations enhanced the binding of ArcR to its target. Using cat fusions, various deletion and substitution mutations within the putative ArcR footprint were shown to cause dramatic reductions in expression from the arcA promoter in vivo, confirming that the 27-bp sequence is required for optimal expression and induction of the ADS by arginine. Mutation of two putative catabolite response elements (CREs) within the arc promoter region showed that both CREs contribute to catabolite repression. A thorough understanding of the regulation of the ADS in S. gordonii and related organisms is needed to develop ways to exploit arginine catabolism for the control of oral diseases. Identification of the ArcR and CcpA binding sites lays the foundation for a more complete understanding of the complex interactions of multiple regulatory proteins with elements in the arc promoter region.
Zeng
Lin
L
Department of Oral Biology, University of Florida, 1600 S.W. Archer Rd., Gainesville, FL 32610-0424, USA.
Dong
Yiqian
Y
Burne
Robert A
RA
D004269:DNA, Bacterial / Q000032:analysis; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D006867:Hydrolases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid / Q000502:physiology*; D013291:Streptococcus / Q000201:enzymology / Q000235:genetics*
D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D006867:Hydrolases; C019723:arginine deiminase