Anaerobic growth promotes synthesis of colonization factors encoded at the Vibrio pathogenicity island in Vibrio cholerae El Tor

Periplasmic proteins encoded by VCA0261–0260 and VC2216 genes together with copA and cueR products are required for copper tolerance but not for virulence in Vibrio cholerae

“…Peptides corresponding to VCA0260 and VCA0261 proteins were identified from a single protein spot, suggesting that both ORFs coded for a single product. In addition, the predicted gene products of these two ORFs were similar to the N-and C-halves of a single putative copper-binding protein in Vibrio parahaemolyticus (Marrero et al, 2009) and to the copper-tolerance protein Cot in Pseudomonas fluorescens DF57 (Tom-Petersen et al, 2001), respectively. Since the cot homologue PA2807 is the most upregulated gene in copper-adapted Pseudomonas aeruginosa PAO1 and its disruption leads to a slight reduction in copper tolerance (Teitzel et al, 2006), we decided to study copper homeostasis in V. cholerae.…”

“…Copper sensitivity was determined on syncase medium, to be consistent with proteomic experiments carried out previously (Marrero et al, 2009). A 20 ml aliquot of a 10 26 dilution from exponentially grown cells of the wild-type or each mutant strain was spread on syncase plates containing increasing concentrations of copper.…”

“…V. cholerae O1 El Tor C7258 and N16961 strains were used for genetic manipulations. Bacterial strains were grown at 37 uC in Luria-Bertani (LB) broth or in syncase medium (Marrero et al, 2009). When necessary, ampicillin and IPTG were added at 100 mg ml 21 and 0.1 mM, respectively.…”

“…On: Thu, 10 May 2018 11:30:17 fingerprinting and the MS/MS ion search option of the MASCOT program, as previously described (Marrero et al, 2009). Additionally, gel strips containing the recombinant polypeptide were crushed and used to immunize New Zealand white rabbits, following standard procedures, to produce specific antisera.…”

“…The survival systems used by the vibrios inside and outside the human host are relevant scientific issues and their disclosure has been facilitated by the availability of the complete genome sequence of this bacterium (Heidelberg et al, 2000), together with improved methodologies for systems biology studies. We previously examined the effect of aerobiosis, anaerobiosis and microaerobiosis on the proteome of V. cholerae by 2D gel electrophoresis (combining IEF and SDS-PAGE) and MS analyses (Marrero et al, 2009). One relevant finding was that anaerobic growth promoted the synthesis of colonization factors encoded by the vibrio pathogenicity island in V. cholerae El Tor.…”