1887

Abstract

Enterohaemorrhagic (EHEC) colonizes and proliferates at the mucosal surface, inducing severe diarrhoea. Short-chain fatty acids (SCFAs) are abundant in the intestine owing to the metabolic activity of microflora, and are important for colonic health. We found that, although a high concentration of SCFAs inhibited the growth of EHEC, at low concentrations, the SCFAs markedly enhanced the expression of the virulence genes required for cell adherence and the induction of attaching and effacing (A/E) lesions. Of the SCFAs tested, butyrate markedly enhanced the expression of these virulence-associated genes, even at the low concentration of 1.25 mM, but acetate and propionate showed only a small effect at concentrations higher than 40 mM. Butyrate enhanced the promoter activity of the operon, which encodes a global regulator of the LEE genes, Ler. This enhancement was dependent on a regulator, PchA. Butyrate sensing was completely abrogated by the deletion of , the gene for the leucine-responsive regulatory protein, Lrp. Expression of a constitutively active mutant of Lrp enhanced the expression of the LEE genes in the absence of butyrate, and a response-defective Lrp derivative reduced the response to butyrate. Thus, upon entering the distal ileum, EHEC may respond to the higher butyrate level via Lrp by increasing its virulence expression, leading to efficient colonization of the target niche.

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2009-02-01
2019-10-21
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Effect of each SCFA on growth. Bacteria were grown overnight in LB and diluted 100-fold with DMEM containing 0.1 M MOPS (pH 6.7) and 0, 5, 10, 20, 40, or 80 mM SCFA or NaCl. [ PDF] (562 kb) Amount of EspB and Tir in bacteria grown with DMEM containing 0.1 M MOPS (pH 6.7) (lane 1), and 20 mM sodium glutamate (lane 2), 20 mM NaCl (lane 3), 20 mM sodium acetate (lane 4), 20 mM sodium propionate (lane 5), or 20 mM sodium butyrate (lane 6). [ PDF] (609 kb) Amount of Lrp in bacteria grown with DMEM containing 20 mM NaCl or SCFA, determined by immunoblotting with Lrp-specific antiserum. [ PDF] (455 kb) Binding of Lrp to the promoter region but not to the or regulatory region. EHEC -FLAG was grown in DMEM containing 20 mM sodium butyrate or NaCl to mid-exponential phase at 37 C. The cultures were used for ChIP analysis with anti-FLAG antibody (as described by Abe , 2008). DNA fragments corresponding to the regulatory region of (P ) or (P ) or (P ) were detected by PCR from input DNA (DNA in supernatant) or precipitated without antibody (ChIP DNA without antibody) or precipitated with anti-FLAG antibody (ChIP DNA with anti-FLAG). The PCR products were separated on agarose gel and detected after staining with ethidium bromide. PlivJ DNA, PpchA DNA, and PLEE1 DNA correspond to 4316147-4316624, 1183578-1183927, and 4620275-4620724 of EHEC O157 Sakai chromosome, respectively. [ PDF] (444 kb) Global regulation by horizontally transferred regulators establishes the pathogenicity of Escherichia coli. , 25-38.

PDF

Effect of each SCFA on growth. Bacteria were grown overnight in LB and diluted 100-fold with DMEM containing 0.1 M MOPS (pH 6.7) and 0, 5, 10, 20, 40, or 80 mM SCFA or NaCl. [ PDF] (562 kb) Amount of EspB and Tir in bacteria grown with DMEM containing 0.1 M MOPS (pH 6.7) (lane 1), and 20 mM sodium glutamate (lane 2), 20 mM NaCl (lane 3), 20 mM sodium acetate (lane 4), 20 mM sodium propionate (lane 5), or 20 mM sodium butyrate (lane 6). [ PDF] (609 kb) Amount of Lrp in bacteria grown with DMEM containing 20 mM NaCl or SCFA, determined by immunoblotting with Lrp-specific antiserum. [ PDF] (455 kb) Binding of Lrp to the promoter region but not to the or regulatory region. EHEC -FLAG was grown in DMEM containing 20 mM sodium butyrate or NaCl to mid-exponential phase at 37 C. The cultures were used for ChIP analysis with anti-FLAG antibody (as described by Abe , 2008). DNA fragments corresponding to the regulatory region of (P ) or (P ) or (P ) were detected by PCR from input DNA (DNA in supernatant) or precipitated without antibody (ChIP DNA without antibody) or precipitated with anti-FLAG antibody (ChIP DNA with anti-FLAG). The PCR products were separated on agarose gel and detected after staining with ethidium bromide. PlivJ DNA, PpchA DNA, and PLEE1 DNA correspond to 4316147-4316624, 1183578-1183927, and 4620275-4620724 of EHEC O157 Sakai chromosome, respectively. [ PDF] (444 kb) Global regulation by horizontally transferred regulators establishes the pathogenicity of Escherichia coli. , 25-38.

PDF

Effect of each SCFA on growth. Bacteria were grown overnight in LB and diluted 100-fold with DMEM containing 0.1 M MOPS (pH 6.7) and 0, 5, 10, 20, 40, or 80 mM SCFA or NaCl. [ PDF] (562 kb) Amount of EspB and Tir in bacteria grown with DMEM containing 0.1 M MOPS (pH 6.7) (lane 1), and 20 mM sodium glutamate (lane 2), 20 mM NaCl (lane 3), 20 mM sodium acetate (lane 4), 20 mM sodium propionate (lane 5), or 20 mM sodium butyrate (lane 6). [ PDF] (609 kb) Amount of Lrp in bacteria grown with DMEM containing 20 mM NaCl or SCFA, determined by immunoblotting with Lrp-specific antiserum. [ PDF] (455 kb) Binding of Lrp to the promoter region but not to the or regulatory region. EHEC -FLAG was grown in DMEM containing 20 mM sodium butyrate or NaCl to mid-exponential phase at 37 C. The cultures were used for ChIP analysis with anti-FLAG antibody (as described by Abe , 2008). DNA fragments corresponding to the regulatory region of (P ) or (P ) or (P ) were detected by PCR from input DNA (DNA in supernatant) or precipitated without antibody (ChIP DNA without antibody) or precipitated with anti-FLAG antibody (ChIP DNA with anti-FLAG). The PCR products were separated on agarose gel and detected after staining with ethidium bromide. PlivJ DNA, PpchA DNA, and PLEE1 DNA correspond to 4316147-4316624, 1183578-1183927, and 4620275-4620724 of EHEC O157 Sakai chromosome, respectively. [ PDF] (444 kb) Global regulation by horizontally transferred regulators establishes the pathogenicity of Escherichia coli. , 25-38.

PDF

Effect of each SCFA on growth. Bacteria were grown overnight in LB and diluted 100-fold with DMEM containing 0.1 M MOPS (pH 6.7) and 0, 5, 10, 20, 40, or 80 mM SCFA or NaCl. [ PDF] (562 kb) Amount of EspB and Tir in bacteria grown with DMEM containing 0.1 M MOPS (pH 6.7) (lane 1), and 20 mM sodium glutamate (lane 2), 20 mM NaCl (lane 3), 20 mM sodium acetate (lane 4), 20 mM sodium propionate (lane 5), or 20 mM sodium butyrate (lane 6). [ PDF] (609 kb) Amount of Lrp in bacteria grown with DMEM containing 20 mM NaCl or SCFA, determined by immunoblotting with Lrp-specific antiserum. [ PDF] (455 kb) Binding of Lrp to the promoter region but not to the or regulatory region. EHEC -FLAG was grown in DMEM containing 20 mM sodium butyrate or NaCl to mid-exponential phase at 37 C. The cultures were used for ChIP analysis with anti-FLAG antibody (as described by Abe , 2008). DNA fragments corresponding to the regulatory region of (P ) or (P ) or (P ) were detected by PCR from input DNA (DNA in supernatant) or precipitated without antibody (ChIP DNA without antibody) or precipitated with anti-FLAG antibody (ChIP DNA with anti-FLAG). The PCR products were separated on agarose gel and detected after staining with ethidium bromide. PlivJ DNA, PpchA DNA, and PLEE1 DNA correspond to 4316147-4316624, 1183578-1183927, and 4620275-4620724 of EHEC O157 Sakai chromosome, respectively. [ PDF] (444 kb) Global regulation by horizontally transferred regulators establishes the pathogenicity of Escherichia coli. , 25-38.

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