The transcription activator GvpE of halophilic Archaea and its interaction partners GvpD, TBP and TFB

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The action of two gene regulators, the transcription activator GvpE, and GvpD exhibiting a repressing function during gas vesicle formation of haloarchaea, will be investigated at the promoter and the posttranscriptional level. The two oppositely oriented promoters PA and PD of the gvp gene cluster are separated by 35 nucleotides, and the 20-nt UAS elements required for GvpE-mediated activation of both promoters overlap with their distal portions in the centre. The simultaneous GvpE-mediated activation of both promoters will be studied by quantitative real-time PCR in wild-type and promoter mutants. The GvpE-responsive element locates next to BRE and TATA-box, and GvpE contacts the TATA-box binding protein TBP. We will investigate whether GvpE also contacts the RNA polymerase or TFB in Halobacterium salinarum. The contact sites of GvpE and TBP, and also the interaction of GvpE with the repressor protein GvpD (that leads to the degradation of GvpE) will be investigated to derive a model for these protein:protein interactions during the regulation. We will monitor the expression of the different tfb and tbp genes under different growth conditions to determine their possible role in the regulation of haloarchaeal gene expression.