Pyrimidine regulation of tandem promoters for carAB in Salmonella typhimurium

Abstract

The carAB operon of Salmonella typhimurium encodes the two subunits of the enzyme carbamoylphosphate synthetase. Transcription of the operon is initiated at tandem promoters that are subject to control by pyrimidines and arginine. Pyrimidine regulation was examined by quantitative primer extension experiments under conditions in which densitometric measurements of the transcripts were linear with the amount of RNA. RNA was obtained from mutant strains that permit manipulations of pyrimidine nucleotide pools. The data showed that a uridine nucleotide repressed the upstream promoter (Pl), whereas arginine repressed the downstream promoter (P2). Exogenous cytidine, which increased the intracellular CTP pool in certain mutant strains, did not affect either promoter. However, CTP limitation resulted in derepression of the pyrimidine-specific promoter as well as the downstream arginine-specific promoter. The effect of pyrimidines on P2 was confirmed in a carA::lacZ transcriptional fusion in which the activity of the pyrimidine-specific promoter was abolished. Primer extension experiments with an argR::Tn10 derivative showed that repression of Pl by uridine nucleotides did not require a functional arginine repressor and that repression of P2 by arginine did not interfere with elongation of transcripts initiated at the upstream Pl promoter.