title: Diverse roles for HspR in Campylobacter jejuni revealed by the proteome, transcriptome and phenotypic characterization of an hspR mutant.
abstract: Campylobacter jejuni is a leading cause of bacterial gastroenteritis in the developed world. The role of a homologue of the negative transcriptional regulatory protein HspR, which in other organisms participates in the control of the heat-shock response, was investigated. Following inactivation of hspR in C. jejuni, members of the HspR regulon were identified by DNA microarray transcript profiling. In agreement with the predicted role of HspR as a negative regulator of genes involved in the heat-shock response, it was observed that the transcript amounts of 13 genes were increased in the hspR mutant, including the chaperone genes dnaK, grpE and clpB, and a gene encoding the heat-shock regulator HrcA. Proteomic analysis also revealed increased synthesis of the heat-shock proteins DnaK, GrpE, GroEL and GroES in the absence of HspR. The altered expression of chaperones was accompanied by heat sensitivity, as the hspR mutant was unable to form colonies at 44 degrees C. Surprisingly, transcriptome analysis also revealed a group of 17 genes with lower transcript levels in the hspR mutant. Of these, eight were predicted to be involved in the formation of the flagella apparatus, and the decreased expression is likely to be responsible for the reduced motility and ability to autoagglutinate that was observed for hspR mutant cells. Electron micrographs showed that mutant cells were spiral-shaped and carried intact flagella, but were elongated compared to wild-type cells. The inactivation of hspR also reduced the ability of Campylobacter to adhere to and invade human epithelial INT-407 cells in vitro, possibly as a consequence of the reduced motility or lower expression of the flagellar export apparatus in hspR mutant cells. It was concluded that, in C. jejuni, HspR influences the expression of several genes that are likely to have an impact on the ability of the bacterium to successfully survive in food products and subsequently infect the consumer.
authors.affiliation: Department of Veterinary Pathobiology, The Royal Veterinary and Agricultural University, Stigbøjlen 4, DK-1958 Frederiksberg C, Denmark.
authors.forename: Marianne Thorup
authors.initials: MT
authors.lastname: Andersen
authors.affiliation: Department of Veterinary Pathobiology, The Royal Veterinary and Agricultural University, Stigbøjlen 4, DK-1958 Frederiksberg C, Denmark.
authors.forename: Lone
authors.initials: L
authors.lastname: Brøndsted
authors.affiliation: Institute of Food Research, Colney, Norwich NR4 7UA, UK.
authors.forename: Bruce M
authors.initials: BM
authors.lastname: Pearson
authors.affiliation: Institute of Food Research, Colney, Norwich NR4 7UA, UK.
authors.forename: Francis
authors.initials: F
authors.lastname: Mulholland
authors.affiliation: Institute of Food Research, Colney, Norwich NR4 7UA, UK.
authors.forename: Mary
authors.initials: M
authors.lastname: Parker
authors.affiliation: Institute of Food Research, Colney, Norwich NR4 7UA, UK.
authors.forename: Carmen
authors.initials: C
authors.lastname: Pin
authors.affiliation: Institute of Food Research, Colney, Norwich NR4 7UA, UK.
authors.forename: Jerry M
authors.initials: JM
authors.lastname: Wells
authors.affiliation: Department of Veterinary Pathobiology, The Royal Veterinary and Agricultural University, Stigbøjlen 4, DK-1958 Frederiksberg C, Denmark.
authors.forename: Hanne
authors.initials: H
authors.lastname: Ingmer
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D016123:Campylobacter jejuni / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity / Q000502:physiology*; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D018869:Heat-Shock Response; D006801:Humans; D009068:Movement; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010641:Phenotype; D020543:Proteome; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D006360:Heat-Shock Proteins; C108444:HspR protein, bacteria; D020543:Proteome; D012097:Repressor Proteins