Hook proteins in membrane trafficking & neurogeneration

描述（由申请人提供）： 神经退行性疾病，如亨廷顿病、肌萎缩侧索硬化症、 硬化症或帕金森病有一个共同的特点， 错误折叠蛋白质的积累。当错误折叠的蛋白质在神经元中积累时 它们不是均匀分布的。相反，它们集中在包容性 尸体这些包涵体是如何与 神经退行性疾病还不太清楚。 其中一类包涵体，即包涵体，在微管中形成 组织中心在一个积极的过程，需要微管为基础的 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。钩蛋白构成了 卷曲螺旋蛋白家族，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 在这项拨款中，我们将联合收割机结合果蝇的遗传学方法，细胞生物学方法， 哺乳动物组织培养细胞和生物化学实验方法 在体外表征Hook蛋白的共有功能，以及 Hook2在错误折叠蛋白质的细胞运输中的特定作用。 在规范目标1中，我们将确定微管结合的相关性， 使用体外生物化学方法和遗传学方法的组合， 果蝇的实验在此背景下，我们还将探讨 Hook蛋白与细胞质动力蛋白和 动力菌素 在规范目标2中，我们将表征Hook蛋白与不同的免疫球蛋白的结合。 细胞器，并确定介导这些相互作用的受体。 在Spec.Aim 3中，我们将确定Hook 2在形成 攻击体和使用显性负性形式的Hook2 操纵不同错误折叠蛋白质的聚集。 在Spec.Aim 4中，我们将确定Hook蛋白的结构域， 它们在神经元中的极化分布以及Hook蛋白在 在大鼠海马神经元中建立神经元极性。

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.