Dual control by purines and pyrimidines of the expression of the pyrD gene of Salmonella typhimurium

Authors


Correspondence to: R.A. Kelln, Department of Chemistry, University of Regina, Regina, Saskatchewan, Canada, S4S OA2.

Abstract

Abstract Expression of the Salmonella typhimurium pyrD gene was found to be repressed two-fold when cells were grown in the presence of hypoxanthine. Purine-mediated repression was evident for reporter plasmids containing pyrD-lacZ transcriptional or translational fusions, indicating that regulation was being exercised at the level of transcriptional initiation. In a strain harbouring a purR6::Tn10 mutation inactivating the purine regulon repressor (PurR), expression of pyrD was not repressed by hypoxanthine. Gel retardation experiments provided evidence that PurR binds to a PUR box centered 27 base pairs upstream of the upstream of the −35 region of the pyrD promoter. Site-directed mutagenesis was used to decrease the similarity of the putative PUR box to the consensus sequence; each mutation eliminated binding of PurR to the mutated DNA in vitro and abolished repression by hypoxanthine in purR+ cells in vivo. Regulation by pyrimidines was unaffected by either of the two PUR box mutations, showing that purine and pyrimidine control of pyrD expression can operate independently.

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