disulfide bond formation and toxr activity in vibrio cholerae二硫键的形成和霍乱弧菌toxr活动.pdfVIP

Disulfide Bond Formation and ToxR Activity in Vibrio cholerae Vera H. I. Fengler, Eva C. Boritsch, Sarah Tutz, Andrea Seper, Hanna Ebner, Sandro Roier, Stefan Schild, Joachim Reidl* Institute of Molecular Biosciences, University of Graz, Humboldtstrasse, Graz, Austria Abstract Virulence factor production in Vibrio cholerae is complex, with ToxRS being an important part of the regulatory cascade. Additionally, ToxR is the transcriptional regulator for the genes encoding the major outer membrane porins OmpU and OmpT. ToxR is a transmembrane protein and contains two cysteine residues in the periplasmic domain. This study addresses the influence of the thiol-disulfide oxidoreductase system DsbAB, ToxR cysteine residues and ToxR/ToxS interaction on ToxR activity. The results show that porin production correlates with ToxR intrachain disulfide bond formation, which depends on DsbAB. In contrast, formation of ToxR intrachain or interchain disulfide bonds is dispensable for virulence factor production and in vivo colonization. This study further reveals that in the absence of ToxS, ToxR interchain disulfide bond formation is facilitated, whereat cysteinyl dependent homo- and oligomerization of ToxR is suppressed if ToxS is coexpressed. In summary, new insights into gene regulation by ToxR are presented, demonstrating a mechanism by which ToxR activity is linked to a DsbAB dependent intrachain disulfide bond formation. Citation: Fengler VHI, Boritsch EC, Tutz S, Seper A, Ebner H, et al. (2012) Disulfide Bond Formation and ToxR Activity in Vibrio cholerae. PLoS ONE 7(10): e47756. doi:10.1371/journal.pone.0047756 Editor: Michael Hensel, University of Osnabrueck, Germany Received August 2, 2012; Accepted September 20, 2012; Published October 29, 2012 Copyright: 2012 Fengler et al. This is an open-access article distributed under the terms of the C