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Applied and Environmental Microbiology, November 2007, p. 7367-7372, Vol. 73, No. 22
0099-2240/07/$08.00+0     doi:10.1128/AEM.01497-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Regulation of a Novel Acidithiobacillus caldus Gene Cluster Involved in Metabolism of Reduced Inorganic Sulfur Compounds{triangledown}

Olena I. Rzhepishevska,1 Jorge Valdés,2 Liucija Marcinkeviciene,3 Camelia Algora Gallardo,1 Rolandas Meskys,3 Violaine Bonnefoy,4 David S. Holmes,2 and Mark Dopson1*

Molecular Biology, Umeå University, SE-901 87 Umeå, Sweden,1 Center for Bioinformatics and Genome Biology, Life Science Foundation, MIFAB and Andrés Bello University, Santiago, Chile,2 Department of Molecular Microbiology and Biotechnology, Institute of Biochemistry, Mokslininku 12, Vilnius LT-08662, Lithuania,3 CNRS, IBSM, Laboratoire de Chimie Bactérienne, 31 Chemin J. Aiguier, 13402 Marseille Cedex 20, France4

Received 4 July 2007/ Accepted 7 September 2007

Acidithiobacillus caldus has been proposed to play a role in the oxidation of reduced inorganic sulfur compounds (RISCs) produced in industrial biomining of sulfidic minerals. Here, we describe the regulation of a new cluster containing the gene encoding tetrathionate hydrolase (tetH), a key enzyme in the RISC metabolism of this bacterium. The cluster contains five cotranscribed genes, ISac1, rsrR, rsrS, tetH, and doxD, coding for a transposase, a two-component response regulator (RsrR and RsrS), tetrathionate hydrolase, and DoxD, respectively. As shown by quantitative PCR, rsrR, tetH, and doxD are upregulated to different degrees in the presence of tetrathionate. Western blot analysis also indicates upregulation of TetH in the presence of tetrathionate, thiosulfate, and pyrite. The tetH cluster is predicted to have two promoters, both of which are functional in Escherichia coli and one of which was mapped by primer extension. A pyrrolo-quinoline quinone binding domain in TetH was predicted by bioinformatic analysis, and the presence of an o-quinone moiety was experimentally verified, suggesting a mechanism for tetrathionate oxidation.

* Corresponding author. Mailing address: Molecular Biology, Umeå University, SE-901 87 Umeå, Sweden. Phone: 46 (0)90 7856769. Fax: 46 (0)90 772630. E-mail: mark.dopson{at}molbiol.umu.se

{triangledown} Published ahead of print on 14 September 2007.

Applied and Environmental Microbiology, November 2007, p. 7367-7372, Vol. 73, No. 22
0099-2240/07/$08.00+0     doi:10.1128/AEM.01497-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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