ArgR-dependent repression of arginine and histidine transport genes in Escherichia coli K-12.
In Escherichia coli L-arginine is taken up by three periplasmic binding protein-dependent transport systems that are encoded by two genetic loci: the artPIQM-artJ and argT-hisJQMP gene clusters. The transcription of the artJ, artPIQM and hisJQMP genes and operons is repressed by liganded ArgR, whereas argT, encoding the LAO (lysine, arginine, ornithine) periplasmic binding protein, is insensitive to the repressor. Here we characterize the repressible Esigma70 P artJ, P artP and P hisJ promoters and demonstrate that the cognate operators consist of two 18 bp ARG boxes separated by 3 bp. Determination of the energy landscape of the ArgR-operator contacts by missing contact probing and mutant studies indicated that each box of a pair contributes to complex formation in vitro and to the repressibility in vivo, but to a different extent. The organization of the ARG boxes and promoter elements in the control regions of the uptake genes is distinct from that of the arginine biosynthetic genes. The hisJQMP operon is the first member of the E. coli ArgR regulon, directly repressed by liganded ArgR, where none of the core promoter elements overlaps the ARG boxes. Single round in vitro transcription assays and DNase I footprinting experiments indicate that liganded ArgR inhibits P artJ and P artP promoter activity by steric exclusion of the RNA polymerase. In contrast, ArgR-mediated repression of P hisJ by inhibition of RNA polymerase binding appears to occur through topological changes of the promoter region.
Caldara
Marina
M
Erfelijkheidsleer en Microbiologie, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium.
Minh
Phu Nguyen Le
PN
Bostoen
Sophie
S
Massant
Jan
J
Charlier
Daniel
D
D026905:Amino Acid Transport Systems / Q000235:genetics*; D026923:Amino Acid Transport Systems, Basic / Q000235:genetics / Q000378:metabolism; D001120:Arginine / Q000378:metabolism*; D001483:Base Sequence; D048168:Escherichia coli K12 / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D005798:Genes, Bacterial; D006639:Histidine / Q000378:metabolism*; D008954:Models, Biological; D008969:Molecular Sequence Data; D009154:Mutation; D009875:Operator Regions, Genetic; D033902:Periplasmic Binding Proteins / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000378:metabolism*; D014158:Transcription, Genetic
D026905:Amino Acid Transport Systems; D026923:Amino Acid Transport Systems, Basic; C490177:ArgR protein, E coli; C097523:ArtJ protein, E coli; D029968:Escherichia coli Proteins; D033902:Periplasmic Binding Proteins; D012097:Repressor Proteins; C018963:histidine-binding protein; D006639:Histidine; D001120:Arginine