Characterization of the PAS domain in the sensor-kinase BvgS: mechanical role in signal transmission

BMC Microbiology

Research Article

Elian Dupré¹  Françoise Jacob-Dubuisson¹  Camille Locht¹  Rudy Antoine¹  Julien Herrou²  Alexandre Wohlkonig³ 

[1] Institut Pasteur de Lille, Center for Infection and Immunity, Lille, France;CNRS UMR8204, Lille, France;INSERM U1019, Lille, France;Univ Lille Nord de France, 59019, Lille, France;Institut Pasteur de Lille, Center for Infection and Immunity, Lille, France;CNRS UMR8204, Lille, France;INSERM U1019, Lille, France;Univ Lille Nord de France, 59019, Lille, France;Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL, USA;VIB Department of Structural Biology, Free University of Brussels, Pleinlaan 2, 1050, Brussels, Belgium;

关键词: Two-component system;    Bordetella;    Virulence regulation;    PAS domain;    Signaling;   

DOI  :  10.1186/1471-2180-13-172

 received in 2013-04-17, accepted in 2013-07-19,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundIn bacteria, signal-transduction two-component systems are major players for adaptation to environmental stimuli. The perception of a chemical or physical signal by a sensor-kinase triggers its autophosphorylation. The phosphoryl group is then transferred to the cognate response regulator, which mediates the appropriate adaptive response. Virulence of the whooping cough agent Bordetella pertussis is controlled by the two-component system BvgAS. Atypically, the sensor-kinase BvgS is active without specific stimuli at 37°C in laboratory conditions and is inactivated by the addition of negative chemical modulators. The structure of BvgS is complex, with two tandem periplasmic Venus flytrap domains and a cytoplasmic PAS domain that precedes the kinase domain, which is followed by additional phosphotransfer domains. PAS domains are small, ubiquitous sensing or regulatory domains. The function of the PAS domain in BvgS remains unknown.ResultsWe showed that recombinant BvgS PAS proteins form dimers that are stabilized by α helical regions flanking the PAS core. A structural model of the PAS domain dimer was built and probed by site-directed mutagenesis and by biochemical and functional analyses. Although we found no ligands for the PAS domain cavity, its integrity is required for signaling. We also showed that the structural stability of the PAS core and its proper coupling to its flanking N- and C-terminal α helices are crucial for BvgS activity.ConclusionsWe propose that a major function of the BvgS PAS domain is to maintain conformational signals arising from mechanical strain generated by the periplasmic domain. The tight structure of the PAS core and its connections with the upstream and downstream helices ensure signaling to the kinase domain, which determines BvgS activity. Many mild substitutions that map to the PAS domain keep BvgS active but make it unresponsive to negative modulators, supporting that modulation increases conformational strain in the protein.

【 授权许可】

Unknown   
© Dupré et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.