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

转录激活子GvpE和GvpD这两个基因调控因子在嗜盐古菌气泡形成过程中的作用将从启动子和转录后水平进行研究。GVP基因簇的两个相反方向的启动子PA和PD被35个核苷酸隔开，GvpE介导的两个启动子激活所需的20个核苷酸UAS元件与它们在中心的远端部分重叠。GvpE介导的两个启动子的同时激活将通过实时定量聚合酶链式反应在野生型和启动子突变体中进行研究。GvpE反应元件位于BRE和TATA-box旁边，GvpE与TATA-box结合蛋白TBP接触。我们将调查GvpE是否也接触盐生杆菌中的RNA聚合酶或TFB。将研究GvpE和TBP的接触部位，以及GvpE与阻遏蛋白GvpD的相互作用(导致GvpE的降解)，以推导出这些蛋白质的模型：调节过程中的蛋白质相互作用。我们将监测不同的tfb和tbp基因在不同生长条件下的表达，以确定它们在调控嗜盐古生菌基因表达中的可能作用。

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.