INSULIN RECEPTOR GENE AND HORMONE SPECIFIC RESPONSE

计划中的研究使用了最近分离的人类胰岛素受体 研究跨膜信号转导的机制和途径 多型性胰岛素反应可通过与胰岛素结合而产生 感受器。通过明智地改变受体的cdna，包括 与同源和异源受体的结构域交换，形成 截短受体与突变和随后的表达 异源细胞，我们建议确定结构要求 胰岛素的特异性反应包括结合、内化和 各种胰岛素特有的功能。我们将搜索细胞内的 在过度产生受体的细胞中的反应的效应者。这个 跨膜信号转导的结构基础将通过使用 针对分子不同区域的单抗组合，如 探测和物理方法，如果可能的话。我们将测试它的作用 酪氨酸激酶结构域AS的细胞定位和诱变 细胞转化的原因。将使用IR cDNAs序列 作为一个探针来分离和表征感兴趣的独特区域 IR基因及其在多种细胞中表达调控的研究 类型。与报告分子相连的IR调节区的重组构建 功能将被用来测试控制IR基因的基因区域 表情。将尝试分析IR中可能存在的缺陷 人类群体中的结构或调节。最后，使用方法 与分离IR基因所需的类似，我们建议 胰岛素样生长因子-I受体的分离、鉴定及比较 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.