THYROTROPIN-RELEASING HORMONE RECEPTOR MOLECULAR BIOLOGY

这项研究计划的广泛，长期目标是确定 促甲状腺激素释放激素（TRH）的作用机制，以便更好地 了解细胞调节的机制，通过所有的细胞外信号 分子，如激素、神经递质和生长因子， 一些病理过程会扰乱细胞调节。 具体而言是 垂体前叶细胞TRH受体的分子生物学将是 研究了 TRH受体的互补DNA（cDNA）具有非常接近的 被克隆了 克隆策略包括表达功能性TRH 小鼠垂体促甲状腺激素受体（TtT） 非洲爪蟾卵母细胞肿瘤。 我们首先建议对cDNA进行测序， 然后用它和推导的受体蛋白的氨基酸序列， 研究确定TRH受体的组织分布， TRH受体的结构-功能关系，特别是 TRH受体关系的分子细节，特别是 其与TRH相互作用及其与鸟嘌呤偶联的分子细节 核苷酸结合调节（G）蛋白，以及调节机制 TRH受体数量，这是一个被证明的机制，调节TRH 行动上 研究将使用GH 3大鼠脑垂体和大鼠垂体进行。 组织中 采用双脱氧核苷酸法的测序策略已被 设计的 将使用放射性标记的cDNA进行原位杂交 probes. 将进行cDNA的选择性突变，然后用于 转化GH-Y细胞，GH 3的亚克隆，不含TRH受体或可检测到 受体mRNA活性，以确定TRH的功能结构域 受体的 并且，TRH受体数量的调节机制将是 通过测量的水平和合成和降解率的研究， 受体蛋白和mRNA，以及mRNA活性的互补测量 在非洲爪蟾卵母细胞，以确定是否有变化， 翻译效率 TRH受体的磷酸化将是 研究其可能的调节机制。

The broad, long term objective of this research program is to define the mechanism of action of thyrotropin-releasing hormone (TRH) so as to better understand the mechanism of cell regulation by all extracellular signal molecules, such as hormones, neurotransmitters and growth factors and of some pathologic processes that disorder cell regulation. Specifically, the molecular biology of the TRH receptor from anterior pituitary cells will be studied. A complementary DNA (cDNA) for the TRH receptor has very nearly been cloned. The cloning strategy involved expression of a functional TRH receptor from a mouse pituitary thyroid-stimulating hormone-producing (TtT) tumor in Xenopus laevis oocytes. We first propose to sequence the cDNA and then use it and the deduced amino acid sequence of the receptor protein in studies to define the tissue distribution of the TRH receptor, the structure-function relations of the TRH receptor, in particular the molecular details of its relations of the TRH receptor, in particular the molecular details of its interaction with TRH and its coupling to a guanine nucleotide binding regulatory (G) protein, and the mechanism of modulation of TRH receptor number, which is a proven mechanism for regulation of TRH action. Studies will be performed using GH3 rat pituitary glands and rat tissues. A sequencing stategy using the dideoxynucleotide method has been devised. In situ hybridization will be performed with radiolabeled cDNA probes. Selective mutations of the cDNA will be made and then used to transform GH-Y cells, a subclone of GH3 without TRH receptors or detectable receptor mRNA activity, to define the functional domains of the TRH receptor. And, the mechanism of regulation of TRH receptor number will be studied by measuring the levels and synthesis and degradation rates of the receptor protein and mRNA, with complementary measurements of mRNA activity made in Xenopus oocytes to determine whether there are changes in translational efficiency. Phosphorylation of the TRH receptor will be studied as a possible mechanism of its regulation.