Purification and functional characterization of PecS, a regulator of virulence‐factor synthesis in <i>Erwinia chrysanthemi</i>

Praillet, Thierry; Nasser, William; Robert‐Baudouy, Janine; Reverchon, Sylvie

doi:10.1111/j.1365-2958.1996.tb02626.x

Cited by 67 publications

(79 citation statements)

References 40 publications

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“…The expression of kdgM is 3-fold repressed with the presence of glucose in the medium or in a crp mutant, indicating that kdgM is submitted to the catabolite repression via the CRP activator. A 7-fold increase in kdgM expression was observed in a pecS mutant (40,20). This regulation by PecS seems to occur essentially during the late exponential or early stationary phase of growth (data not shown).…”

Section: Kdgm Is a Major Outer Membrane Protein Of Pectinolyticmentioning

confidence: 89%

“…Band shift assays were performed with the KdgR-, CRP-, and PecS-purified proteins (32,33,40). The labeled DNA fragment (50,000 cpm) and 5-100 nM of the purified regulator were incubated for 30 min at 30°C in 20 l of binding buffer, and the reaction mixtures were submitted to electrophoresis on a 4% non-denaturing polyacrylamide gel, as described previously (33,40).…”

Section: Methodsmentioning

confidence: 99%

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The Oligogalacturonate-specific Porin KdgM of Erwinia chrysanthemi Belongs to a New Porin Family

Blot¹,

Berrier²,

Hugouvieux‐Cotte‐Pattat³

et al. 2002

Journal of Biological Chemistry

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The phytopathogenic Gram-negative bacteria Erwinia chrysanthemi secretes pectinases, which are able to degrade the pectic polymers of plant cell walls, and uses the degradation products as a carbon source for growth. We characterized a major outer membrane protein, KdgM, whose synthesis is strongly induced in the presence of pectic derivatives. The corresponding gene was characterized. Analysis of transcriptional fusions showed that the kdgM expression is controlled by the general repressor of pectinolytic genes, KdgR, by the repressor of hexuronate catabolism genes, ExuR, by the pectinase gene repressor, PecS, and by catabolite repression via the cyclic AMP receptor protein (CRP) transcriptional activator. A kdgM mutant is unable to grow on oligogalacturonides longer than trimers, and its virulence is affected. Electrophysiological experiments with planar lipid bilayers showed that KdgM behaves like a voltage-dependent porin that is slightly selective for anions and that exhibits fast block in the presence of trigalacturonate. In contrast to most porins, KdgM seems to be monomeric. KdgM has no homology with currently known porins, but proteins similar to KdgM are present in several bacteria. Therefore, these proteins might constitute a new family of porin channels.

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Section: Kdgm Is a Major Outer Membrane Protein Of Pectinolyticmentioning

confidence: 89%

Section: Methodsmentioning

confidence: 99%

The Oligogalacturonate-specific Porin KdgM of Erwinia chrysanthemi Belongs to a New Porin Family

Blot¹,

Berrier²,

Hugouvieux‐Cotte‐Pattat³

et al. 2002

Journal of Biological Chemistry

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“…A similar inverted repeat lies between the promoter and start codon of SlyA (see Ludwig et al, 1995). In contrast, the Hpr-binding sites appear to be a somewhat shorter and imperfect inverted repeat (Kallio et al, 1991), whilst the PecS-binding sites appear to be much longer (Praillet et al, 1996) …”

Section: Cinr Binds To Sequences Between Cinr and Cinbmentioning

confidence: 99%

Expression of a Butyrivibrio fibrisolvens E14 gene (cinB) encoding an enzyme with cinnamoyl ester hydrolase activity is negatively regulated by the product of an adjacent gene (cinR)

Dalrymple

Swadling

1997

Microbiology

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A second cinnamoyl ester hydrolase (CEH) encoding gene (cinB) has been characterized from the ruminal bacterium Butyrivibrio fibrisolvens E14. CinB is more similar to CinA (previously named Cinl) (28% amino acid identity), the first CEH described from B. fibrisolvens E14, than either of the enzymes are to any other member of the family of hydrolases to which they belong. Upstream of cinB, and in the opposite orientation, is a gene (cinR) encoding a protein with substantial similarity to members of the MarR family of negative regulators of bacterial gene expression. By alignment of these sequences, a possible helix-turn-helix DNA-binding domain has been identified. CinR was expressed at a high level in Escherichia coli using the lac promoter. In E. coli CinR repressed the expression of CinB, but had no effect on the expression of CinA. In gel mobility-shift assays, CinR bound specifically to the cinR-cinB intergenic region. Two identical 16 nucleotide inverted repeats adjacent to the putative PcinR and PcinB promoters are likely binding sites for CinR. The addition of FAXX (O-[5-O-(trans-feruloyl)-α-L-arabinofuranosyl]-(1,3)-O-ß-D-xylopyranosyl-(1,4)-D-xylopyranose) and Fara [5-O-(trans-feruloyl)-arabinofuranose], but not xylobiose, ferulic acid and a number of other soluble components of hemicellulose, inhibited the binding of CinR to DNA.

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“…In E. chrysanthemi, it has been demonstrated that the full expression of the pectin catabolism genes requires the presence of the cAMP-CRP complex Reverchon et al, 1997) and that the KdgR repressor essentially mediates the induction of these catabolic pathway genes by pectic compounds (Reverchon et al, 1991;Nasser et al, 1994). In addition to kdgR, two other loci involved in negative regulation of the pectinase gene expression, pecS-pecM and pecT, were also characterized in E. chrysanthemi, but the signal to which they respond still remains unknown (Praillet et al, 1996;Reverchon et al, 1994;Surgey et al, 1996;Castillo and Reverchon, 1997).…”

Section: Introductionmentioning

confidence: 99%

Characterization of the Erwinia chrysanthemi expI–expR locus directing the synthesis of two N‐acyl‐homoserine lactone signal molecules

Nasser

Bouillant

Salmond

et al. 1998

Molecular Microbiology

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168

120

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SummaryThe plant pathogen Erwinia chrysanthemi produces three acyl-homoserine lactones (acyl-HSLs). One has been identified as N-(3-oxohexanoyl)-homoserine lactone (OHHL), and the two others were supposed to be N-(hexanoyl)-homoserine lactone (HHL) and N-(decanoyl)-homoserine lactone (DHL). The genes for a quorum-sensing signal generator (expI ) and a response regulator (expR ) were cloned. These genes are convergently transcribed and display high similarity to the expI-expR genes of Erwinia carotovora. ExpI is responsible for both OHHL and HHL production. Inactivation of expI had little effect on pectinase synthesis in E. chrysanthemi, as expression of only two of the pectate lyase genes, pelA and pelB, was decreased. E. chrysanthemi expR mutants still produced acyl-HSL and pectinases. However, gel shift and DNAse I footprinting experiments showed that the purified E. chrysanthemi ExpR protein binds specifically to the promoter regions of the five major pel genes. Addition of OHHL modified the ExpR-DNA bandshift profiles, indicating that ExpR interacts with OHHL and binds to DNA in different ways, depending on the OHHL concentration. Localization of the ExpR binding sites just upstream of promoter regions suggests that ExpR functions as an activator of pel expression in the presence of OHHL. The absence of a phenotype in expR mutants strongly suggests that at least an additional interchangeable ExpR homologue exists in E. chrysanthemi. Finally, transcription of expI ::uidA and expR ::uidA fusions is dependent on the population density, suggesting the existence of a quorum-sensing hierarchy in E. chrysanthemi. These results suggest that the expI-expR locus is part of a complex autoregulatory system that controls quorum sensing in E. chrysanthemi.

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Purification and functional characterization of PecS, a regulator of virulence‐factor synthesis in Erwinia chrysanthemi

Cited by 67 publications

References 40 publications

The Oligogalacturonate-specific Porin KdgM of Erwinia chrysanthemi Belongs to a New Porin Family

The Oligogalacturonate-specific Porin KdgM of Erwinia chrysanthemi Belongs to a New Porin Family

Expression of a Butyrivibrio fibrisolvens E14 gene (cinB) encoding an enzyme with cinnamoyl ester hydrolase activity is negatively regulated by the product of an adjacent gene (cinR)

Characterization of the Erwinia chrysanthemi expI–expR locus directing the synthesis of two N‐acyl‐homoserine lactone signal molecules

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