The downstream regulatory element of the proU operon of Salmonella typhimurium inhibits open complex formation by RNA polymerase at a distance.
The intracellular concentration of K(+)-glutamate, chromatin-associated proteins, and a downstream regulatory element (DRE) overlapping with the coding sequence, have been implicated in the regulation of the proU operon of Salmonella typhimurium. The basal expression of the proU operon is low, but it is rapidly induced when the bacteria are grown in media of high osmolarity (e.g. 0.3 M NaCl). It has previously been suggested that increased intracellular concentrations of K(+)-glutamate activate the proU promoter in response to increased extracellular osmolarity. We show here that the activation of the proU promoter by K(+)-glutamate in vitro is nonspecific, and the in vivo regulation cannot simply be mimicked in vitro. In vivo specificity requires both the chromatin-associated protein H-NS and the DRE; they are both needed to maintain repression of proU expression at low osmolarity. How H-NS and the DRE repress the proU promoter in vivo has so far been unclear. We show that, in vivo, the DRE acts at a distance to inhibit open complex formation at the proU promoter.
Jordi
B J
BJ
Department of Bacteriology, Institute of Infectious Diseases and Immunology, Faculty of Veterinary Sciences, Yalelaan 1, 3508 TD Utrecht, The Netherlands. B.Jordi@vet.uu.nl
Higgins
C F
CF
D001426:Bacterial Proteins / Q000235:genetics*; D018983:DNA Footprinting; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D009876:Operon; D009994:Osmolar Concentration; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000235:genetics*; D012486:Salmonella typhimurium / Q000235:genetics*; D037941:Silencer Elements, Transcriptional; D001618:beta-Lactamases / Q000235:genetics
D001426:Bacterial Proteins; D012097:Repressor Proteins; C095098:proU silencer; D012321:DNA-Directed RNA Polymerases; D001618:beta-Lactamases