Regulation of virulence by butyrate sensing in enterohaemorrhagic Escherichia coli, by N. Nakanishi, K. Tashiro, S. Kuhara, T. Hayashi, N. Sugimoto and T. Tobe
Microbiology vol. 155, part 2, pp. 521 - 530
Fig. S1. 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)
Fig. S2. 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)
Fig. S3. Amount of Lrp in bacteria grown with DMEM containing 20 mM NaCl or SCFA, determined by immunoblotting with Lrp-specific antiserum. [PDF] (455 kb)
Fig. S4. Binding of Lrp to the livJ promoter region but not to the pchA or LEE1 regulatory region. EHEC lrp-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 et al., 2008). DNA fragments corresponding to the regulatory region of livJ (PlivJ) or pchA (PpchA) or LEE1 (PLEE1) 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)
REFERENCE
Abe, H., Miyahara, A., Oshima, T., Tashiro, K., Ogura, Y., Kuhara, S., Ogasawara, N., Hayashi, T. & Tobe, T. (2008). Global regulation by horizontally transferred regulators establishes the pathogenicity of Escherichia coli. DNA Res 15, 25-38.
Copyright © 2009 Society for General Microbiology.
