title: CadA negatively regulates Escherichia coli O157:H7 adherence and intestinal colonization.
abstract: Adherence of pathogenic Escherichia coli strains to intestinal epithelia is essential for infection. For enterohemorrhagic E. coli (EHEC) serotype O157:H7, we have previously demonstrated that multiple factors govern this pathogen's adherence to HeLa cells (A. G. Torres and J. B. Kaper, Infect. Immun. 71:4985-4995, 2003). One of these factors is CadA, a lysine decarboxylase, and this protein has been proposed to negatively regulate virulence in several enteric pathogens. In the case of EHEC strains, CadA modulates expression of the intimin, an outer membrane adhesin involved in pathogenesis. Here, we inactivated cadA in O157:H7 strain 86-24 to investigate the role of this gene in EHEC adhesion to tissue-cultured monolayers, global gene expression patterns, and colonization of the infant rabbit intestine. The cadA mutant did not possess lysine decarboxylation activity and was hyperadherent to tissue-cultured cells. Adherence of the cadA mutant was nearly twofold greater than that of the wild type, and the adherence phenotype was independent of pH, lysine, or cadaverine in the media. Additionally, complementation of the cadA defect reduced adherence back to wild-type levels, and it was found that the mutation affected the expression of the intimin protein. Disruption of the eae gene (intimin-encoding gene) in the cadA mutant significantly reduced its adherence to tissue-cultured cells. However, adherence of the cadA eae double mutant was greater than that of an 86-24 eae mutant, suggesting that the enhanced adherence of the cadA mutant is not entirely attributable to enhanced expression of intimin in this background. Gene array analysis revealed that the cadA mutation significantly altered EHEC gene expression patterns; expression of 1,332 genes was downregulated and that of 132 genes was upregulated in the mutant compared to the wild-type strain. Interestingly, the gene expression variation shows an EHEC-biased gene alteration including intergenic regions. Two putative adhesins, flagella and F9 fimbria, were upregulated in the cadA mutant, suggestive of their association with adherence in the absence of the Cad regulatory mechanism. In the infant rabbit model, the cadA mutant outcompeted the wild-type strain in the ileum but not in the cecum or mid-colon, raising the possibility that CadA negatively regulates EHEC pathogenicity in a tissue-specific fashion.
authors.affiliation: Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555-1070, USA.
authors.forename: Roberto C
authors.initials: RC
authors.lastname: Vazquez-Juarez
authors.forename: Jeeba A
authors.initials: JA
authors.lastname: Kuriakose
authors.forename: David A
authors.initials: DA
authors.lastname: Rasko
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Ritchie
authors.forename: Melissa M
authors.initials: MM
authors.lastname: Kendall
authors.forename: Terry M
authors.initials: TM
authors.lastname: Slater
authors.forename: Mala
authors.initials: M
authors.lastname: Sinha
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Luxon
authors.forename: Vsevolod L
authors.initials: VL
authors.lastname: Popov
authors.forename: Matthew K
authors.initials: MK
authors.lastname: Waldor
authors.forename: Vanessa
authors.initials: V
authors.lastname: Sperandio
authors.forename: Alfredo G
authors.initials: AG
authors.lastname: Torres
mesh_terms: D018829:Adhesins, Bacterial / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D001422:Bacterial Adhesion / Q000502:physiology*; D015153:Blotting, Western; D002262:Carboxy-Lyases / Q000235:genetics / Q000378:metabolism*; D004927:Escherichia coli Infections / Q000235:genetics / Q000378:metabolism* / Q000382:microbiology; D019453:Escherichia coli O157 / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000235:genetics; D015870:Gene Expression; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D020411:Oligonucleotide Array Sequence Analysis; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D018829:Adhesins, Bacterial; D029968:Escherichia coli Proteins; C073819:eaeA protein, E coli; D002262:Carboxy-Lyases; C021520:lysine decarboxylase
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