Journal Article

Characterization of a two-component signal transduction system that controls arsenite oxidation in the chemolithoautotroph NT-26

Sunita Sardiwal, Joanne M. Santini, Thomas H. Osborne and Snezana Djordjevic

in FEMS Microbiology Letters

Volume 313, issue 1, pages 20-28
Published in print December 2010 |
Published online November 2010 | e-ISSN: 1574-6968 | DOI: http://dx.doi.org/10.1111/j.1574-6968.2010.02121.x

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Abstract

NT-26 is a chemolithoautotrophic arsenite oxidizer. Understanding the mechanisms of arsenite signalling, tolerance and oxidation by NT-26 will have significant implications for its use in bioremediation and arsenite sensing. We have identified the histidine kinase (AroS) and the cognate response regulator (AroR) involved in the arsenite-dependent transcriptional regulation of the arsenite oxidase aroBA operon. AroS contains a single periplasmic sensory domain that is linked through transmembrane helices to the HAMP domain that transmits the signal to the kinase core of the protein. AroR belongs to a family of AAA+ transcription regulators that interact with DNA through a helix-turn-helix domain. The presence of the AAA+ domain as well as the RNA polymerase σ54-interaction sequence motif suggests that this protein regulates transcription through interaction with RNA polymerase in a σ54-dependent fashion. The kinase core of AroS and the receiver domain of AroR were heterologously expressed and purified and their autophosphorylation and transphosphorylation activities were confirmed. Using site-directed mutagenesis, we have identified the phosphorylation sites on both proteins. Mutational analysis in NT-26 confirmed that both proteins are essential for arsenite oxidation and the AroS mutant affected growth with arsenite, also implicating it in the regulation of arsenite tolerance. Lastly, arsenite sensing does not appear to involve thiol chemistry.

Keywords: histidine kinase; response regulator; AAA+ protein; arsenite sensing; arsenite oxidation

Journal Article.  5557 words.  Illustrated.

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