Hook proteins in membrane trafficking & neurogeneration

描述(由申请人提供)： 亨廷顿病、肌萎缩侧索硬化症等神经退行性疾病 硬化症或帕金森氏症有一个共同的特征，即慢性病 错误折叠的蛋白质堆积。随着错误折叠的蛋白质在神经元中积累 它们的分布并不均匀。相反，它们集中在包容中。 身体。这些包涵体是如何与肿瘤进展相联系的 神经退行性疾病还没有得到很好的了解。 在微管上形成一类包涵体，即侵袭体。 需要基于微管的主动过程中的组织中心 运输。我们最近发现，活性浓度的错误折叠 侵袭体中的蛋白质涉及HOOK2蛋白。钩蛋白构成一种 卷曲卷曲蛋白家族，与微管结合并影响 哺乳动物细胞和果蝇中不同细胞器的组织。在……里面 这笔拨款，我们将结合果蝇的遗传方法，细胞生物学 哺乳动物组织培养细胞和生化实验的研究进展 体外鉴定Hook蛋白的共同功能，以及 HOOK2在错误折叠蛋白的细胞运输中的特殊作用。 在等级库目的1，我们将确定钩的微管结合的相关性 利用体外和遗传生物化学方法相结合的蛋白质 在果蝇身上的实验。在此背景下，我们还将探索 Hook蛋白与胞质动力蛋白与胞质动力蛋白复合体的相互作用 动力蛋白。 在等级库目的2，我们将表征Hook蛋白与不同的 细胞器，并确定介导这些相互作用的受体。 在等级库目标3，我们将确定HOOK2在HOK2的形成中的作用 攻击体和使用显性否定形式HOOK2的潜力 操纵不同错误折叠的蛋白质的聚集。 在等级库目的4，我们将确定Hook蛋白的结构域负责 它们在神经元中的极化分布和Hook蛋白在神经元中的作用 建立大鼠海马神经元的神经元极性。

DESCRIPTION (provided by applicant): Neurodegenerative diseases such as Huntington's disease, amyotrophic lateral sclerosis or Parkinson's disease share one common feature, the slow accumulation of misfolded proteins. As misfolded proteins accumulate in neurons they are not evenly distributed. Instead, they are concentrated in inclusion bodies. How these inclusion bodies are linked to the progression of neurodegenerative diseases is not well understood. One class of inclusion bodies, aggresomes, are formed at the microtubule organizing center in an active process that requires microtubule-based transport. We recently discovered that the active concentration of misfolded proteins in aggresomes involves the Hook2 protein. Hook proteins constitute a family of coiled-coil proteins which bind to microtubules and affect the organization of different organelles in mammalian cells and in Drosophila. In this grant, we will combine genetic approaches in Drosophila, cell biological approaches in mammalian tissue culture cells and biochemical experiments in-vitro to characterize shared functions of Hook proteins, as well as the specific role of Hook2 in the cellular trafficking of misfolded proteins. In Spec. Aim 1, we will determine the relevance of microtubule binding of Hook proteins using a combination of biochemical approaches in vitro and genetic experiments in Drosophila. In this context we will also explore the potential interaction of Hook proteins with the complex between cytoplasmic Dynein and Dynactin. In Spec. Aim 2, we will characterize the binding of Hook proteins to different organelles and identify the receptors that mediate these interactions. In Spec. Aim 3, we will determine the role of Hook2 in the formation of aggresomes and the potential of using dominant-negative forms of Hook2 to manipulate the aggregation of different misfolded proteins. In Spec. Aim 4, we will determine the domains of Hook proteins responsible for their polarized distribution in neurons and the role of Hook proteins in establishing neuronal polarity in rat hippocampal neurons.