The RcsC sensor kinase is required for normal biofilm formation in <i>Escherichia coli</i> K‐12 and controls the expression of a regulon in response to growth on a solid surface

Ferrières, Lionel; Clarke, David J.

doi:10.1046/j.1365-2958.2003.03815.x

Cited by 218 publications

(226 citation statements)

References 68 publications

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“…Nevertheless, the introduction of a mutation that blocked colanic acid production, gmd : : lplacMu53 (Ferrières & Clarke, 2003), into the rcsC : : Tn10 background had no effect on biofilm formation, confirming that the promiscuous expression of the cps operon was not responsible for the biofilm defect of this strain ( Supplementary Fig. S2).…”

Section: Mutations In Rcsc and Rcsd Affect Biofilm Formationmentioning

confidence: 78%

Elevated levels of σ S inhibit biofilm formation in Escherichia coli: a role for the Rcs phosphorelay

2009

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The Rcs phosphorelay is composed of RcsC, RcsD and the response regulator RcsB, and this signalling pathway has been implicated in virulence and biofilm formation in many enteric bacteria. It was previously shown that a mutation in rcsC resulted in defective biofilm formation in Escherichia coli [Ferrières, L. & Clarke, D. J. (2003) Mol Microbiol 50, 1665–1682]. To identify the molecular mechanisms underlying the observed biofilm defect we carried out a screen looking for suppressor mutants that restored biofilm formation in the rcsC mutant background. One of the mutants was identified to be in rprA, a gene encoding a small RNA molecule that is involved in the post-transcriptional control of the alternative sigma factor, σ S. The expression of rprA is regulated by the Rcs phosphorelay, and there are elevated σ S levels present in the rcsC mutant due to the overexpression of rprA in this background. Using different approaches, we have established that the increase in σ S levels is responsible for the biofilm defect. Therefore, the Rcs phosphorelay is involved in maintaining appropriate levels of σ S during biofilm formation in E. coli.

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Section: Mutations In Rcsc and Rcsd Affect Biofilm Formationmentioning

confidence: 78%

Elevated levels of σ S inhibit biofilm formation in Escherichia coli: a role for the Rcs phosphorelay

2009

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“…A recent genome-wide analysis suggests that the Rcs regulon of E. coli may include more than 40 loci (Hagiwara et al, 2003;Oshima et al, 2002). Another study shows that the RcsC sensor kinase is required for normal biofilm development in E. coli and that more than 150 genes are under Rcs control (Ferrieres & Clarke, 2003).…”

Section: Discussionmentioning

confidence: 99%

Virulence attenuation in Salmonella enterica rcsC mutants with constitutive activation of the Rcs system

et al. 2005

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Mutations in rcsC that result in constitutive colanic acid capsule synthesis were obtained in Salmonella enterica serovar Typhimurium. Most rcsC alleles were dominant; however, recessive rcsC alleles were also found, in agreement with the postulated double role (positive and negative) of RcsC on the activation of the RcsB/C phosphorelay system. Salmonella rcsC mutants with constitutive activation of the Rcs system are severely attenuated for virulence in BALB/c mice and their degree of attenuation correlates with the level of Rcs activation. Partial relief of attenuation by a gmm mutation indicates that capsule overproduction is one of the factors leading to avirulence in constitutively activated rcsC mutants. INTRODUCTIONThe Rcs phosphorelay system was initially characterized in Escherichia coli as a two-component positive regulator of colanic acid capsule synthesis encoded by rcsC and rcsB (Brill et al., 1988;Gottesman et al., 1985;Stout & Gottesman, 1990). The sensor protein RcsC, a hybrid histidine kinase, and the transcriptional activator RcsB are the main components of the system, which also includes a second transcriptional activator, RcsA (Stout et al., 1991) and an intermediate phosphotransmitter, YojN (Takeda et al., 2001). The Rcs system is also a negative regulator of the flagellar master operon flhDC (Francez-Charlot et al., 2003), whose products are necessary for the expression of genes involved in flagellum synthesis, motility and chemotaxis.The first environmental signal shown to strongly and rapidly induce colanic acid synthesis was osmotic upshift (Sledjeski & Gottesman, 1996). This effect was mediated by the Rcs system. More recently it has been shown that the Rcs phosphorelay can be activated when wild-type cells are grown at 20 uC in the presence of glucose and a high concentration of Zn 2+ (Hagiwara et al., 2003). Furthermore, several genetic conditions that result in alterations in envelope composition or integrity are known to activate the Rcs system. For example, a mutation in the mdoH gene involved in the biosynthesis of membrane-derived oligosaccharides (Ebel et al., 1997) Here, we describe the isolation of Salmonella rcsC mutants with constitutive activation of the Rcs system, and the characterization of amino acid substitutions that lead to different degrees of activation of the system. Virulence trials with constitutively activated rcsC mutants provide a direct demonstration of the link between activation of the Rcs system and avirulence in mice. We also show that virulence attenuation associated with activation of the RcsB/C system is partially due to colanic acid overproduction. METHODSBacterial strains, bacteriophages and strain construction.S. enterica serovar Typhimurium strains used in this study are described in Table 1. Unless otherwise indicated, the strains derive from the mouse-virulent strain 14028. Transductional crosses using phage P22 HT 105/1 int201 (Schmieger, 1972) were used for strain construction operations. The transduction protocol was described by Maloy (199...

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“…A survey of gene expression in Pseudomonas aeruginosa biofilms using DNA microarrays has identified tolA as a gene activated in biofilms (Whiteley et al, 2001). In addition, the expression of both tolQRA and csgDEFG is modulated by RcsC (Clavel et al, 1996;Ferrieres & Clarke, 2003), suggesting a potential link between Tol-Pal, Rcs and curli expression. This led us to investigate the potential roles of the tol genes in biofilm formation in laboratory and clinical isolates of E. coli, and of the Rcs system in the regulation of curli synthesis.…”

Section: Introductionmentioning

confidence: 99%

“…The OmpR/EnvZ (Prigent-Combaret et al, 2001;Romling et al, 1998) and CpxR/A (Dorel et al, 1999;Otto & Silhavy, 2002;Prigent-Combaret et al, 2001) phosphorelays also control curli expression. Using microarray analysis, the RcsC sensor kinase of the RcsB/CD phosphorelay has recently also been found to control csgD expression (Ferrieres & Clarke, 2003 The RcsB/CD His-Asp phosphorelay was initially identified as a positive regulator of the capsular exopolysaccharide biosynthesis gene cluster (wza-wca) in E. coli in association with RcsA (Gottesman & Stout, 1991;Stout, 1994). The response regulator RcsB is activated upon the transfer of a phosphate group from its cognate sensor, RcsC, via a histidine-containing phosphotransmitter domain protein, RcsD.…”

Section: Introductionmentioning

confidence: 99%

Escherichia coli tol and rcs genes participate in the complex network affecting curli synthesis

et al. 2005

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Curli are necessary for the adherence of Escherichia coli to surfaces, and to each other, during biofilm formation, and the csgBA and csgDEFG operons are both required for their synthesis. A recent survey of gene expression in Pseudomonas aeruginosa biofilms has identified tolA as a gene activated in biofilms. The tol genes play a fundamental role in maintaining the outer-membrane integrity of Gram-negative bacteria. RcsC, the sensor of the RcsBCD phosphorelay, is involved, together with RcsA, in colanic acid capsule synthesis, and also modulates the expression of tolQRA and csgDEFG. In addition, the RcsBCD phosphorelay is activated in tol mutants or when Tol proteins are overexpressed. These results led the authors to investigate the role of the tol genes in biofilm formation in laboratory and clinical isolates of E. coli. It was shown that the adherence of cells was lowered in the tol mutants. This could be the result of a drastic decrease in the expression of the csgBA operon, even though the expression of csgDEFG was slightly increased under such conditions. It was also shown that the Rcs system negatively controls the expression of the two csg operons in an RcsA-dependent manner. In the tol mutants, activation of csgDEFG occurred via OmpR and was dominant upon repression by RcsB and RcsA, while these two regulatory proteins repressed csgBA through a dominant effect on the activator protein CsgD, thus affecting curli synthesis. The results demonstrate that the Rcs system, previously known to control the synthesis of the capsule and the flagella, is an additional component involved in the regulation of curli. Furthermore, it is shown that the defect in cell motility observed in the tol mutants depends on RcsB and RcsA. INTRODUCTIONThe ability of bacteria to recognize and adhere to a specific surface is a fundamental aspect of microbial ecology and pathogenesis. Bacterial adhesins and fimbriae confer specific recognition and adhesion to diverse target molecules, such as mammalian host tissue components or inorganic materials. Curli are highly adhesive proteinaceous fibres involved in this process that are produced by Escherichia coli (Vidal et al., 1998) and Salmonella (Collinson et al., 1991). The genes necessary for curli formation are clustered in the csgBA and csgDEFG operons. CsgA encodes the curlin subunit, CsgB is thought to nucleate CsgA curlin fibres (Bian & Normark, 1997), CsgD is a transcriptional activator of the csgBA operon, and CsgE, CsgF and CsgG are three putative curli assembly factors. The lipoprotein CsgG localizes to the inner leaflet of the outer membrane and may serve as a curli assembly platform (Loferer et al., 1997); CsgE and CsgF are thought to be chaperone-like proteins (Chapman et al., 2002). The csg genes appear to be expressed in environmental strains, clinical isolates and some laboratory strains of E. coli.Regulation of curli synthesis is carried out through a complex network of interactions between the transcription factors and the csg regulatory region. Nine regulators have ...

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The RcsC sensor kinase is required for normal biofilm formation in Escherichia coli K‐12 and controls the expression of a regulon in response to growth on a solid surface

Cited by 218 publications

References 68 publications

Elevated levels of σ S inhibit biofilm formation in Escherichia coli: a role for the Rcs phosphorelay

Elevated levels of σ S inhibit biofilm formation in Escherichia coli: a role for the Rcs phosphorelay

Virulence attenuation in Salmonella enterica rcsC mutants with constitutive activation of the Rcs system

Escherichia coli tol and rcs genes participate in the complex network affecting curli synthesis

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