THYROTROPIN-RELEASING HORMONE RECEPTOR MOLECULAR BIOLOGY

这项研究计划的广泛、长期目标是定义 促甲状腺激素释放激素(TRH)的作用机制 了解所有细胞外信号对细胞的调控机制 分子，如激素、神经递质和生长因子以及 一些扰乱细胞调节的病理过程。具体地说， 来自前垂体细胞的TRH受体的分子生物学将是 学习。TRH受体的互补DNA(CDNAs)与 都被克隆了。克隆策略涉及到功能性TRH的表达 小鼠垂体促甲状腺激素(TTT)受体 非洲爪哇卵母细胞肿瘤。我们首先提出了对该基因进行测序的建议 然后用它和推导出的受体蛋白的氨基酸序列在 定义TRH受体的组织分布的研究， 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 核苷酸结合调节蛋白及其调控机制 TRH受体数目，这是一种已被证实的调节TRH的机制 行动。研究将使用GH3大鼠脑垂体腺和大鼠 纸巾。一种使用双脱氧核苷酸方法的测序策略已经 精心设计的。将用放射性标记的cDNAs进行原位杂交 探测器。将对cdna进行选择性突变，然后用于 转化GH-Y细胞，无TRH受体或可检测到的GH3的亚克隆 受体mRNA活性，以确定TRH的功能结构域 受体。并且，调节TRH受体数量的机制将是 通过测量水平、合成和降解率来研究 受体蛋白和信使核糖核酸，以及信使核糖核酸活性的互补测量 在非洲爪哇卵母细胞中制作以确定是否有变化 翻译效率。TRH受体的磷酸化将是 作为一种可能的调节机制进行了研究。

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.