1887

Abstract

GB17 requires thiosulfate for induction of the sulfur-oxidizing (Sox) enzyme system. The genes are divergently oriented to the genes. predicts a transcriptional regulator of the ArsR family and a periplasmic thioredoxin. The homogenote mutant GBΩS carrying a disruption of by the Ω-kanamycin-resistance-encoding interposon expressed a low thiosulfate-oxidizing activity under heterotrophic and mixotrophic growth conditions. This activity was repressed by complementation with , suggesting that SoxR acts as a repressor and SoxS is essential for full expression. Sequence analysis uncovered operator characteristics in the intergenic regions and . In each region a transcription start site was identified by primer extension analysis. Both regions were cloned into the vector pRI1 and transferred to . Strains harbouring pRI1 with or expressed the genes under heterotrophic conditions at a low rate, indicating repressor titration. Sequence analysis of SoxR suggested a helix–turn–helix (HTH) motif at position 87–108 and uncovered an invariant Cys-80 and a cysteine residue at the C-terminus. SoxR was overproduced in with an N-terminal His-tag and purified to near homogeneity. Electrophoretic gel mobility shift assays with SoxR retarded the region as a single band while the region revealed at least two protein–DNA complexes. These data demonstrated binding of SoxR to the relevant DNA. This is believed to be the first report of regulation of chemotrophic sulfur oxidation at the molecular level.

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2005-05-01
2020-07-13
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