title: rpoS function is essential for bgl silencing caused by C-terminally truncated H-NS in Escherichia coli.
abstract: From evolutionary and physiological viewpoints, the Escherichia coli bgl operon is intriguing because its expression is silent (Bgl(-) phenotype), at least under several laboratory conditions. H-NS, a nucleoid protein, is known as a DNA-binding protein involved in bgl silencing. However, we previously found that bgl expression is still silent in a certain subset of hns mutations, each of which results in a defect in its DNA-binding ability. Based on this fact, we proposed a model in which a postulated DNA-binding protein(s) has an adapter function by interacting with both the cis-acting element of the bgl promoter and the mutated H-NS. To identify such a presumed adapter molecule, we attempted to isolate mutants exhibiting the Bgl(+) phenotype in the background of hns60, encoding the mutant H-NS protein lacking the DNA-binding domain by random insertion mutagenesis with the mini-Tn10cam transposon. These isolated mutations were mapped to five loci on the chromosome. Among these loci, three appeared to be leuO, hns, and bglJ, which were previously characterized, while the other two were novel. Genetic analysis revealed that the two insertions are within the rpoS gene and in front of the lrhA gene, respectively. The former encodes the stationary-phase-specific sigma factor, sigma(S), and the latter encodes a LysR-like DNA-binding protein. It was found that sigma(S) is defective in both types of mutant cells. These results showed that the rpoS function is involved in the mechanism underlying bgl silencing, at least in the hns60 background used in this study. We also examined whether the H-NS homolog StpA has such an adapter function, as was previously proposed. Our results did not support the idea that StpA has an adapter function in the genetic background used.
authors.affiliation: Laboratory of Molecular Microbiology, School of Agriculture, Nagoya University, Chikusa-ku, Nagoya 464-8601, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Ohta
authors.forename: C
authors.initials: C
authors.lastname: Ueguchi
authors.forename: T
authors.initials: T
authors.lastname: Mizuno
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D020868:Gene Silencing; D005960:Glucosides / Q000378:metabolism*; D018832:Molecular Chaperones; D009154:Mutation; D009876:Operon; D010446:Peptide Fragments / Q000378:metabolism; D017384:Sequence Deletion; D012808:Sigma Factor / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D005960:Glucosides; C031908:H-NS protein, bacteria; C490682:LrhA protein, E coli; C095974:LrhA protein, bacteria; D018832:Molecular Chaperones; D010446:Peptide Fragments; D012808:Sigma Factor; C090104:StpA protein, E coli; D014157:Transcription Factors; C069567:sigma factor KatF protein, Bacteria