Characterization of the arginine deiminase operon of Streptococcus rattus FA-1.
The arginine deiminase system (ADS) is of critical importance in oral biofilm pH homeostasis and microbial ecology. The ADS consists of three enzymes. Arginine is hydrolyzed by AD (ArcA) to generate citrulline and ammonia. Citrulline is then converted to ornithine and carbamoylphosphate via ornithine carbamoyltransferase (ArcB). Finally, carbamate kinase (ArcC) transfers a phosphate from carbamoylphosphate to ADP, yielding ATP. Ammonia production from this pathway protects bacteria from lethal acidification, and ATP production provides a source of energy for the cells. The purpose of this study was to initiate a characterization of the arc operon of Streptococcus rattus, the least cariogenic and sole ADS-positive member of the mutans streptococci. Using an arcB gene fragment obtained by degenerate PCRs, the FA-1 arc operon was identified in subgenomic DNA libraries and sequence analysis was performed. Results showed that the genes encoding the AD pathway in S. rattus FA-1 are organized as an arcABCDT-adiR operon gene cluster, including the enzymes of the pathway, an arginine-ornithine antiporter (ArcD) and a putative regulatory protein (AdiR). The arcA transcriptional start site was identified by primer extension, and a sigma(70)-like promoter was mapped 5' to arcA. Reverse transcriptase PCR was used to establish that arcABCDT could be cotranscribed. Reporter gene fusions and AD assays demonstrated that the operon is regulated by substrate induction and catabolite repression, the latter apparently through a CcpA-dependent pathway.
Department of Oral Biology, University of Florida, Gainesville, Florida 32610, USA.
Ann
A
Griswold
Yi-Ywan M
YY
Chen
Jennifer A
JA
Snyder
Robert A
RA
Burne
D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005798:Genes, Bacterial; D017930:Genes, Reporter; D006867:Hydrolases / Q000235:genetics*; D008969:Molecular Sequence Data; D009876:Operon; D013291:Streptococcus / Q000201:enzymology* / Q000235:genetics*; D013295:Streptococcus mutans / Q000201:enzymology / Q000235:genetics
D004269:DNA, Bacterial; D006867:Hydrolases; C019723:arginine deiminase