INSULIN RECEPTOR GENE AND HORMONE SPECIFIC RESPONSE

计划中的研究采用了最近分离的人胰岛素受体 研究跨膜信号传导的机制， 多型胰岛素反应可由胰岛素结合产生， 受体。 通过明智地改变受体cDNA，包括 同源和异源受体的结构域交换， 截短的受体和诱变以及随后在 异源细胞，我们建议确定结构要求， 胰岛素特异性反应包括结合、内化和 各种胰岛素特异性功能。 我们将寻找细胞内的 在过量产生受体的细胞中的反应的效应物。 的 跨膜信号传导的结构基础将通过使用 针对分子各个区域的单克隆抗体组， 探测器和物理方法，如果可能的话。 我们将测试 酪氨酸激酶结构域的细胞定位和诱变， 细胞转化的原因。 将使用IR cDNA序列 作为探针分离和表征感兴趣的独特区域， IR基因，并研究其在多种细胞中的表达调控 类型 与报告基因连接的IR调节区的重组构建体 功能将用于测试控制IR基因的基因区域 表情 将尝试分析IR中可能存在的缺陷 结构或调节。 最后，使用方法 类似于分离IR基因所需的那些，我们提出， 分离并鉴定IGF-I受体， 研究胰岛素受体。

The projected studies employ the recently isolated human insulin receptor to study the mechanism of transmembrane signalling and the means by which the pleiotypic insulin response can be generated from insulin binding to the receptor. By judicious alteration of the receptor cDNA, including domain swaps with homologous and heterologous receptors, formation of truncated receptors and mutagenesis and subsequent expression in heterologous cells, we propose to determine the structural requirements for the insulin specific responses including binding, internalization and the various insulin specific functions. We will search for the intracellular effectors of the response in cells overproducing the receptor. The structural basis of transmembrane signalling will be analyzed by employing panels of monoclonal antibodies to various regions of the molecule as probes and physical methodologies if possible. We will test the role of cellular localization and mutagenesis of the tyrosine kinase domain as causes for cellular transformation. The IR cDNA sequence will be employed as a probe to isolate and characterize the unique regions of interest in the IR gene and study the control of its expression in a variety of cell types. Recombinant constructs of IR regulatory regions linked to reporter functions will be used to test the regions of the gene controlling IR gene expression. Attempts will be made to analyze possible defects in IR structure or regulation in human populations. Finally, using methods similar to those required for isolation of the IR gene we propose to isolate and characterize the IGF-I receptor and carry out comparative studies with the insulin receptor.