Transcription of the Corynebacterium diphtheriae hmuO gene is regulated by iron and heme.
The hmuO gene is required for the utilization of heme and hemoglobin as iron sources by Corynebacterium diphtheriae. The product of hmuO has homology to eukaryotic heme oxygenases which are involved in the degradation of heme and the release of iron. To investigate the mechanism of hmuO regulation, a promoterless lacZ gene present on the promoter-probe vector pCM502 was placed under transcriptional control of the hmuO promoter. In C. diphtheriae C7, optimal expression from the hmuO promoter was obtained only in the presence of heme or hemoglobin under low-iron conditions. Expression of hmuO in high-iron medium containing heme was repressed five- to sixfold from that seen under low-iron conditions in the presence of heme. Transcription from the hmuO promoter in the absence of heme or hemoglobin was fully repressed in high-iron medium and was expressed at very low levels in iron-depleted conditions. Expression studies with tile hmuO-lacZ fusion construct in C7hm723, a dtxR mutant of C7, and in a hmuO mutant of C. diphtheriae HC1 provided further evidence that transcription of the hmuO promoter is repressed by DtxR and iron and activated by heme. In Escherichia coli, the hmuO promoter was expressed at very low levels under all conditions examined. Gel mobility shift assays and DNase I footprinting experiments indicated that DtxR binds in a metal-dependent manner to a sequence that overlaps the putative hmuO promoter. Total cellular RNA isolated from C. diphtheriae was used to identify the transcriptional start site for the hmuO gene. Northern blot analysis suggested that the hmuO mRNA was monocistronic and that transcription was heme inducible.
Schmitt
M P
MP
Division of Bacterial Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892, USA. Schmitt@A1.cber.fda.gov
D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D003353:Corynebacterium diphtheriae / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects*; D006418:Heme / Q000494:pharmacology*; D006419:Heme Oxygenase (Decyclizing); D007501:Iron / Q000494:pharmacology*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D006418:Heme; D007501:Iron; D006419:Heme Oxygenase (Decyclizing); C104107:HmuO protein, bacteria