INTERACTIONS OF ANDROGEN RECEPTOR POLYGLUTAMINE TRACT

描述：(申请人摘要)脊髓和延髓肌萎缩症 一种成人起病的神经退行性疾病，以肌肉无力为特征， 萎缩和束状物。它是一个不断壮大的家庭的成员 由CAG扩张引起的神经退行性疾病重复 在相关蛋白编码中编码多聚谷氨酰胺链 序列。我们的假设是在这些细胞中看到的神经退行性变 疾病是由编码蛋白质中的有毒功能的获得引起的 由于聚谷氨酰胺束的存在，这种毒性效应 是通过特定的蛋白质-蛋白质相互作用来调节的。 该项目计划鉴定和分离蛋白质(及其相关的 与含有不同长度的雄激素受体相互作用 多聚谷氨酰胺区域。这将通过两种方式完成。首先，我们将使用 酵母双杂交系统鉴定与氨基相互作用的蛋白质 含有24、45或66个谷氨酰胺的雄激素受体末端区域 重复着。我们还将使用该系统来表征 多聚谷氨酰胺束长度上的相互作用。我们预计这将是 一种相互作用在致病过程中发挥作用，它应该是 在某种程度上受到谷氨酰胺重复长度的影响。第二，我们将 用放射性标记技术筛选λ噬菌体表达文库 分离纯化不同重复长度的雄激素受体蛋白 细菌。这种方法将允许我们对库进行差异筛选 使用含有不同长度的全长雄激素受体 多聚谷氨酰胺区域。该系统还将被开发用于 通过以下任一方法鉴定的蛋白质的结合特性的表征 接近。最后，我们将描述蛋白质的生理学相关性。 通过这些筛选确定的相互作用使用细胞培养和 我们开发的转基因模型系统。 这些实验的结果应该会提供对分子的洞察 扩张的聚谷氨酰胺束对神经元产生毒性的机制， 并可因此提供治疗干预的靶点。在……里面 此外，了解神经元中雄激素受体蛋白的相互作用 将增加我们对类固醇激素受体在 这些细胞。

DESCRIPTION: (Applicant's abstract) Spinal and bulbar muscular atrophy is an adult-onset neurodegenerative disease characterized by muscle weakness, atrophy, and fasciculations. It is a member of a growing family of neurodegenerative diseases caused by the expansion of a CAG repeat that encodes a poly-glutamine tract within the associated protein coding sequence. Our hypothesis is that the neurodegeneration seen in these diseases results from the gain of a toxic function in the encoded protein due to the presence of the poly-glutamine tract, and that this toxic effect is mediated by specific protein-protein interactions. This project proposes to identify and isolate proteins (and their associated cDNAs) that interact with androgen receptor containing different length poly-glutamine tracts. This will be done in two ways. First, we will use a yeast two-hybrid system to identify proteins that interact with the amino terminal region of the androgen receptor containing 24, 45, or 66 glutamine repeats. We will also use this system to characterize the dependence of interactions on the length of the poly-glutamine tract. We expect that for an interaction to play a role in the pathogenic process, it should be affected in some way by the length of the glutamine repeat. Second, we will screen a lambda phage expression library using radioactively-labeled androgen receptor proteins of different repeat lengths purified from bacteria. This method will allow us to differentially screen libraries using full-length androgen receptor containing different length poly-glutamine tracts. This system will also be exploited for characterization of binding properties of proteins identified through either approach. Lastly, we will characterize the physiologic relevance of protein interactions identified through these screens using cell culture and transgenic model systems that we have developed. The results of these experiments should provide insights into the molecular mechanism by which expanded poly-glutamine tracts become toxic to neurons, and may consequently provide targets for therapeutic intervention. In addition, understanding androgen receptor protein interactions in neurons will increase our understanding of the role of steroid hormone receptors in these cells.