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首页> 外文期刊>Molecular and Cellular Biology >Functional coupling of a mammalian somatostatin receptor to the yeast pheromone response pathway.
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Functional coupling of a mammalian somatostatin receptor to the yeast pheromone response pathway.

机译：哺乳动物生长抑素受体与酵母信息素反应途径的功能偶联。

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A detailed analysis of structural and functional aspects of G-protein-coupled receptors, as well as discovery of novel pharmacophores that exert their effects on members of this class of receptors, will be facilitated by development of a yeast-based bioassay. To that end, yeast strains that functionally express the rat somatostatin receptor subtype 2 (SSTR2) were constructed. High-affinity binding sites for somatostatin ([125I-Tyr-11]S-14) comparable to those in native tissues were detected in yeast membrane extracts at levels equivalent to the alpha-mating pheromone receptor (Ste2p). Somatostatin-dependent growth of strains modified by deletion of genes encoding components of the pheromone response pathway was detected through induction of a pheromone-responsive HIS3 reporter gene, enabling cells to grow on medium lacking histidine. Dose-dependent growth responses to S-14 and related SSTR2 subtype-selective agonists that were proportional to the affinity of the ligands for SSTR2 were observed. The growth response required SSTR2, G alpha proteins, and an intact signal transduction pathway. The sensitivity of the bioassay was affected by intracellular levels of the G alpha protein. A mutation in the SST2 gene, which confers supersensitivity to pheromone, was found to significantly enhance the growth response to S-14. In sst2 delta cells, SSTR2 functionally interacted with both a chimeric yeast/mammalian G alpha protein and the yeast G alpha protein, Gpa1p; to promote growth. These yeast strains should serve as a useful in vivo reconstitution system for examination of molecular interactions of the G-protein-coupled receptors and G proteins.
机译：基于酵母的生物测定法的开发将有助于对G蛋白偶联受体的结构和功能方面进行详细分析，以及发现能对此类受体成员发挥作用的新型药效基团。为此，构建了功能上表达大鼠生长抑素受体亚型2（SSTR2）的酵母菌株。在酵母膜提取物中检测到的生长抑素（[125I-Tyr-11] S-14）的高亲和力结合位点与天然组织中的结合位点与α-交配信息素受体（Ste2p）相当。通过诱导信息素应答的HIS3报告基因，检测到通过删除信息素应答途径成分编码基因而修饰的生长抑素依赖性菌株的生长，从而使细胞能够在缺乏组氨酸的培养基上生长。观察到对S-14和相关SSTR2亚型选择性激动剂的剂量依赖性生长反应，该反应与配体对SSTR2的亲和力成比例。生长反应需要SSTR2，G alpha蛋白和完整的信号转导途径。生物测定的敏感性受到细胞内Gα蛋白水平的影响。 SST2基因突变赋予信息素超敏性，被发现可以显着增强对S-14的生长反应。在sst2三角洲细胞中，SSTR2在功能上与嵌合酵母/哺乳动物G alpha蛋白和酵母G alpha蛋白Gpa1p相互作用。促进增长。这些酵母菌株应作为有用的体内重组系统，用于检查G蛋白偶联受体和G蛋白的分子相互作用。

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