Steroid Receptor Regulation by TPR Proteins

TPR 蛋白对类固醇受体的调节

基本信息

批准号：
7003681
负责人：
MICHAEL DAVID CHINKERS
金额：
$ 25.09万
依托单位：
UNIVERSITY OF SOUTH ALABAMA
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-09-15 至 2006-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7003681
关键词：
Steroid Receptor Regulation TPR Proteins

项目摘要

DESCRIPTION (provided by applicant): For proper function, steroid receptors must form complexes with hsp90 and several co-chaperones that bind to the hsp90 C-terminus via TPR domains. The roles of these co-chaperones are poorly understood, but they are likely to regulate receptor localization and binding affinity. We have found differences between TPR, hsp90 binding sites that might be exploited to develop specific inhibitors. The broad, long-term objectives of this proposal are to understand the roles of specific hsp90 co-chaperones in regulating signaling by steroid receptors, and to develop more specific hsp90-targeted drugs than are currently available. Our specific aims are: 1. To determine the roles played by TPR-containing co-chaperones in the glucocorticoid receptor (GR) pathway. We hypothesize that different co-chaperones have some redundant functions but that they also have specific functions resulting from their structural differences. 2. To compare the regulation of different steroid receptors by different TPR-containing hsp90 co-chaperones in living cells. Different steroid receptors associate preferentially with different co-chaperones. We hypothesize that this preferential association is required for normal responses to steroid hormones. 3. To identify peptide inhibitors that specifically block the binding of particular TPR-containing co-chaperones to hsp90. We hypothesize that inhibitors of the binding of particular TPR proteins to hsp90 will differentially alter signaling by different steroid receptors. The health relatedness of the project lies in gaining a better understanding of proteins important in cancer and endocrine disorders, and in obtaining information that could lead to the development of more specific hsp90-targeted drugs than are currently available. The experimental plan is: 1. To use RNAi to test the role of hsp90 co-chaperones in GR nucleocytoplasmic shuttling, formation of hsp90 heterocomplexes, binding affinity, and reporter gene activation. 2. To use RNAi to assess the role of hsp90 co-chaperones in the functions of other steroid receptors. 3. To screen phage display libraries for peptides that specifically block binding of particular co-chaperones to hsp90. To use TAT-fusions and microinjection to test these peptides' effects on steroid receptor function in living cells.

描述（由申请人提供）：对于适当的功能，类固醇受体必须与HSP90和几个通过TPR域结合与HSP90 C末端结合的副群形成复合物。这些共伴侣的作用知之甚少，但它们可能调节受体定位和结合亲和力。我们发现可能利用以开发特定抑制剂的TPR，HSP90结合位点之间的差异。该提案的广泛，长期的目标是了解特定的HSP90共伴侣在调节类固醇受体调节信号传导中的作用，并开发出比目前可用的更具体的HSP90靶向药物。我们的具体目的是：1。确定含TPR的辅助酮在糖皮质激素受体（GR）途径中扮演的角色。我们假设不同的伴侣具有一些冗余功能，但它们也具有由于其结构差异而产生的特定功能。 2。比较活细胞中不同类固醇受体的调节。不同的类固醇受体优先与不同的伴侣相关。我们假设对类固醇激素的正常反应需要这种优先关联。 3。鉴定肽抑制剂，这些肽抑制剂特异性阻断了特定的含TPR的辅助蛋白与HSP90的结合。我们假设特定TPR蛋白与HSP90结合的抑制剂将通过不同类固醇受体的信号差异改变信号。该项目的健康相关性在于对在癌症和内分泌疾病中重要的蛋白质有更好的了解，并获得可能导致比目前可用的更具体的HSP90靶向药物的信息。实验计划是：1。使用RNAi测试Hsp90辅助酮在GR核质shuttling中的作用，HSP90异质复合物的形成，结合亲和力和报告基因激活。 2。使用RNAi评估Hsp90共伴侣在其他类固醇受体功能中的作用。 3。筛选噬菌体显示库的肽，这些肽特异性地阻止了特定的共伴酮与HSP90的结合。使用TAT融合和显微注射来测试这些肽对活细胞类固醇受体功能的影响。