title: Evaluation of the effects of sdiA, a luxR homologue, on adherence and motility of Escherichia coli O157 : H7.
abstract: Quorum-sensing (QS) signalling pathways are important regulatory networks for controlling the expression of genes promoting adherence of enterohaemorrhagic Escherichia coli (EHEC) O157 : H7 to epithelial cells. A recent study has shown that EHEC O157 : H7 encodes a luxR homologue, called sdiA, which upon overexpression reduces the expression of genes encoding flagellar and locus of enterocyte effacement (LEE) proteins, thus negatively impacting on the motility and intimate adherence phenotypes, respectively. Here, we show that the deletion of sdiA from EHEC O157 : H7 strain 86-24, and from a hha (a negative regulator of ler) mutant of this strain, enhanced bacterial adherence to HEp-2 epithelial cells of the sdiA mutant strains relative to the strains containing a wild-type copy of sdiA. Quantitative reverse transcription PCR showed that the expression of LEE-encoded genes ler, espA and eae in strains with the sdiA deletions was not significantly different from that of the strains wild-type for sdiA. Similarly, no additional increases in the expression of LEE genes were observed in a sdiA hha double mutant strain relative to that observed in the hha deletion mutant. While the expression of fliC, which encodes flagellin, was enhanced in the sdiA mutant strain, the expression of fliC was reduced by several fold in the hha mutant strain, irrespective of the presence or absence of sdiA, indicating that the genes sdiA and hha exert opposing effects on the expression of fliC. The strains with deletions in sdiA or hha showed enhanced expression of csgA, encoding curlin of the curli fimbriae, with the expression of csgA highest in the sdiA hha double mutant, suggesting an additive effect of these two gene deletions on the expression of csgA. No significant differences were observed in the expression of the genes lpfA and fimA of the operons encoding long polar and type 1 fimbriae in the sdiA mutant strain. These data indicate that SdiA has no significant effect on the expression of LEE genes, but that it appears to act as a strong repressor of genes encoding flagella and curli fimbriae, and the alleviation of the SdiA-mediated repression of these genes in an EHEC O157 : H7 sdiA mutant strain contributes to enhanced bacterial motility and increased adherence to HEp-2 epithelial cells.
authors.affiliation: Pre-Harvest Food Safety and Enteric Diseases Research Unit, National Animal Disease Center, Ames, IA 50010, USA.
authors.forename: Vijay K
authors.initials: VK
authors.lastname: Sharma
authors.affiliation: Pre-Harvest Food Safety and Enteric Diseases Research Unit, National Animal Disease Center, Ames, IA 50010, USA.
authors.forename: Shawn M D
authors.initials: SMD
authors.lastname: Bearson
authors.affiliation: Agroecosystems Management Research Unit, National Laboratory for Agriculture and the Environment, Agricultural Research Service, United States Department of Agriculture, Ames, IA 50010, USA.
authors.forename: Bradley L
authors.initials: BL
authors.lastname: Bearson
mesh_terms: D001422:Bacterial Adhesion / Q000502:physiology*; D018441:Biofilms; D002460:Cell Line; D003224:Congo Red / Q000378:metabolism; D019453:Escherichia coli O157 / Q000235:genetics / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D010861:Fimbriae, Bacterial / Q000235:genetics; D005407:Flagella / Q000235:genetics; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D009068:Movement; D010750:Phosphoproteins / Q000235:genetics; D012097:Repressor Proteins / Q000737:chemistry; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics / Q000502:physiology*
chemical_list: D029968:Escherichia coli Proteins; C457340:LEE protein, E coli; D010750:Phosphoproteins; D012097:Repressor Proteins; D015534:Trans-Activators; C490806:sdiA protein, E coli; C064778:LuxR autoinducer binding proteins; D003224:Congo Red