title: The mechanism underlying Ler-mediated alleviation of gene repression by H-NS.
abstract: Secretion of effector proteins in Enteropathogeneic Escherichia coli (EPEC) and Enterohemorrhagic Escherichia coli (EHEC) is mediated by a specialized type III secretion system, components of which are encoded in the LEE operons 1 to 5. H-NS, a global repressor in E. coli, silences the expression of LEE operons. Ler, a master regulator in LEE operons, shares 24% amnio acid identity and 44% amino acid similarity to H-NS. Interestingly, rather than a gene silencer, its main role has been characterized as an antagonizing protein that relieves H-NS-mediated transcriptional silencing. In the previous study we reported molecular mechanism for the repression of LEE5 promoter in EPEC and EHEC by H-NS as a protein interaction between upstream DNA-bound H-NS and the αCTD of promoter-bound RNA polymerase. The mechanism underlying Ler-mediated alleviation of the genes repression by H-NS is largely unknown. We examined regulatory effect of these proteins on LEE5p activity using various in vitro tools. Our results revealed that binding affinity of Ler to the LEE5p DNA is about 40 folds greater than that of H-NS as determined by surface plasmon resonance. We verified that Ler binding removed H-NS bound to the same stretch of DNA on LEE5 promoter resulting in a derepression.
authors.affiliation: Department of Microbiology, Kyungpook National University School of Medicine, 680 Gukchaebosang-Ro, Jung-gu, Daegu, 41944, South Korea. Electronic address: shinms@knu.ac.kr.
authors.forename: Minsang
authors.initials: M
authors.lastname: Shin
mesh_terms: D020856:AT Rich Sequence; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D036781:Fimbriae Proteins / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D011401:Promoter Regions, Genetic; D020349:Surface Plasmon Resonance; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D029968:Escherichia coli Proteins; C529458:FimG protein, E coli; C465556:Ler protein, E coli; D015534:Trans-Activators; D036781:Fimbriae Proteins
keywords: Anti-repressor; H-NS; LEE (locus of enterocyte effacement); Ler