Regulation of the Salmonella typhimurium metF gene by the MetR protein.
The metF gene in Escherichia coli and Salmonella typhimurium is under negative transcriptional control by the MetJ repressor. Expression of an S. typhimurium metF-lacZ gene fusion is repressed up to 10-fold by methionine addition to the growth medium in E. coli hosts encoding wild-type MetJ repressor; this repression is not seen in metJ mutants. metR mutations which eliminate the MetR activator protein result in two- to threefold-more-severe repression by the MetJ repressor. In a metJ metR double mutant, however, the level of metF-lacZ expression is the same as in a metJ mutant, suggesting that MetR antagonizes MetJ-mediated methionine repression of the metF promoter. A DNA footprint analysis showed that MetR binds to a DNA fragment carrying the metF promoter and protects two separate regions from DNase I digestion: a 46-bp region from position -50 to -95 upstream of the transcription initiation site and a 24-bp region from about position +62 to +85 downstream of the transcription initiation site and within the metF structural gene. Nucleotide changes in each of the MetR-binding sites away from the consensus sequence disrupt MetR-mediated regulation of the metF-lacZ fusion.
Cowan
J M
JM
Department of Microbiology, University of Iowa, Iowa City 52242.
Urbanowski
M L
ML
Talmi
M
M
Stauffer
G V
GV
D045103:5,10-Methylenetetrahydrofolate Reductase (FADH2); D001426:Bacterial Proteins / Q000378:metabolism*; D001665:Binding Sites; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005800:Genes, Fungal; D007763:Lac Operon; D016297:Mutagenesis, Site-Directed; D010088:Oxidoreductases / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012486:Salmonella typhimurium / Q000201:enzymology / Q000235:genetics*; D015534:Trans-Activators / Q000378:metabolism*
D001426:Bacterial Proteins; C058169:MetR protein, Bacteria; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D010088:Oxidoreductases; D045103:5,10-Methylenetetrahydrofolate Reductase (FADH2)