title: Role of HrcA and CIRCE in the heat shock regulatory network of Bradyrhizobium japonicum.
abstract: A large number of bacteria regulate chaperone gene expression by the CIRCE-HrcA system in which a DNA element called CIRCE serves as binding site for the repressor protein HrcA under non-heat-shock conditions. We have cloned the two consecutive genes hrcA and grpE of Bradyrhizobium japonicum by using a complementation approach that screened for GrpE function. In vivo and in vitro transcript mapping demonstrated that both genes are transcribed separately from RpoH (sigma(32))-dependent promoters. To investigate the supposed negative regulatory function of HrcA, we compared the expression of putative target genes in the wild type with that in an hrcA mutant. Transcription of the CIRCE-associated chaperonin operons groESL(4) and groESL(5), as well as the beta-galactosidase activity derived from corresponding groE-lacZ fusions, was strongly elevated in the hrcA mutant even at physiological temperatures. Expression of other heat shock regulons (RpoH or ROSE dependent) was not affected. To study the activity of HrcA in vitro, we purified a histidine-tagged version of the protein under nondenaturing conditions. Specific binding to the CIRCE element was obtained with a soluble fraction of HrcA in gel retardation experiments.
authors.affiliation: Institut für Mikrobiologie, Eidgenössische Technische Hochschule, CH-8092 Zürich, Switzerland.
authors.forename: A C
authors.initials: AC
authors.lastname: Minder
authors.forename: H M
authors.initials: HM
authors.lastname: Fischer
authors.forename: H
authors.initials: H
authors.lastname: Hennecke
authors.forename: F
authors.initials: F
authors.lastname: Narberhaus
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D020369:Bradyrhizobium / Q000502:physiology*; D018835:Chaperonin 10 / Q000235:genetics; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D018869:Heat-Shock Response / Q000502:physiology*; D008969:Molecular Sequence Data; D010641:Phenotype; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D012097:Repressor Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D012808:Sigma Factor; D013559:Symbiosis / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D001426:Bacterial Proteins; D018835:Chaperonin 10; D004268:DNA-Binding Proteins; C068113:GrpE protein, Bacteria; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; D012097:Repressor Proteins; D012808:Sigma Factor; D014157:Transcription Factors; C069774:heat-shock sigma factor 32