Transcriptional regulation of the uptake [NiFe]hydrogenase genes in Rhodobacter capsulatus.
Transcription of the hupSL genes, which encode the uptake [NiFe]hydrogenase of Rhodobacter capsulatus, is specifically activated by H(2). Three proteins are involved, namely the H(2)-sensor HupUV, the histidine kinase HupT and the transcriptional activator HupR. hupT and hupUV mutants have the same phenotype, i.e. an increased level of hupSL expression (assayed by phupS::lacZ fusion) in the absence of H(2); they negatively control hupSL gene expression. HupT can autophosphorylate its conserved His(217), and in vitro phosphotransfer to Asp(54) of its cognate response regulator, HupR, was demonstrated. The non-phosphorylated form of HupR binds to an enhancer site (5'-TTG-N(5)-CAA) of phupS localized at -162/-152 nt and requires integration host factor to activate fully hupSL transcription. HupUV is an O(2)-insensitive [NiFe]hydrogenase, which interacts with HupT to regulate the phosphorylation state of HupT in response to H(2) availability. The N-terminal domain of HupT, encompassing the PAS domain, is required for interaction with HupUV. This interaction with HupT, leading to the formation of a (HupT)(2)-(HupUV)(2) complex, is weakened in the presence of H(2), but incubation of HupUV with H(2) has no effect on the stability of the heterodimer/tetramer, HupUV-(HupUV)(2), equilibrium. HupSL biosynthesis is also under the control of the global two-component regulatory system RegB/RegA, which controls gene expression in response to redox. RegA binds to a site close to the -35 promoter recognition site and to a site overlapping the integration host factor DNA-binding site (5'-TCACACACCATTG, centred at -87 nt) and acts as a repressor.
Vignais
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PM
Laboratoire de Biochimie et Biophysique des Systèmes Intégrés, UMR 5092 CEA/CNRS/Université Joseph Fourier, Département de Réponse et Dynamique Cellulaires, 17 rue des Martyrs, 38054 Grenoble cedex 9, France. paulette.vignais@cea.fr
Elsen
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Colbeau
A
A
D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D006864:Hydrogenase / Q000235:genetics*; D005810:Multigene Family; D010084:Oxidation-Reduction; D016354:Rhodobacter capsulatus / Q000201:enzymology* / Q000235:genetics; D014158:Transcription, Genetic
C043296:nickel-iron hydrogenase; D006864:Hydrogenase