Molecular Genetics of Cytoplasmic Dyein

描述（由申请人提供）：动力蛋白是一类运动蛋白，参与微管依赖运动的许多方面。特别是细胞质动力蛋白，负责各种形式的囊泡、大分子和有丝分裂运动，以及微管细胞骨架和定向细胞运动的组织。动力蛋白如何发挥作用产生力，以及细胞质动力蛋白如何选择性地针对亚细胞结构的多样性，这是该领域两个主要的突出问题。该提案根据前一个项目期间的进展情况，追求针对每个或这些问题的目标。目的1处理茎，一个15纳米长的投影从马达域延伸，这是负责微管结合。这个Aim将在原子分辨率上确定茎的结构；它将进一步确定其假定的反平行盘绕式d-螺旋内的构象变化，该构象负责微管结合和ATP水解的远程变构耦合。目的II涉及细胞质动力蛋白货物结合的机制。我们最近的研究揭示了ZW10蛋白复合物在将动力蛋白和动力蛋白连接到不同的亚细胞结构中的一般作用。我们将定义负责ZW10膜结合的机制；以高尔基体为重点，我们将确定ZW10相对于Rab6、spectrin和其他与动力蛋白货物结合有关的因子的作用，目的是完整阐明高尔基货物结合途径。Aim III将涉及动力蛋白介导的病毒运动。我们将重点讨论腺病毒作为动力蛋白货物的一种结构简单的致病形式。我们将扩展并完成我们的工作，以确定动力蛋白和腺病毒衣壳之间的相互作用伙伴。总之，这些研究对理解正常和异常细胞和发育生物学的许多方面具有重要意义。最终目的对急性病毒感染的控制和基因传递载体和策略的设计具有特殊意义。

DESCRIPTION (provided by applicant): The dyneins are a class of motor protein involved in many of aspects of microtubule-dependent movement. Cytoplasmic dynein, in particular, is responsible for diverse forms of vesicular, macromolecular, and mitotic movement, as well as organization of the microtubule cytoskeleton and directed cell movement. How the dyneins function to produce force, and how cytoplasmic dynein is selectively targeted to a diversity of subcellular structures represent two of the major outstanding issues in the field. This proposal pursues Aims directed at each or these issues based on progress during the preceding project period. Aim I deals with the stalk, a 15 nm long projection extending from the motor domain, which is responsible for microtubule binding. This Aim will determine the structure of the stalk at atomic resolution; it will further identify conformational changes within its putative antiparallel coiled-coil D-helix responsible for long-range allosteric coupling of microtubule binding and ATP hydrolysis. Aim II addresses mechanisms involved in cytoplasmic dynein cargo binding. Our recent studies have revealed a general role for the ZW10 protein complex in linking dynactin and dynein to diverse subcellular structures. We will define the mechanism responsible for ZW10 membrane binding; focusing on the Golgi apparatus, we will determine the role of ZW10 relative to Rab6, spectrin, and other factors implicated in dynein cargo binding, with the goal of a complete elucidation of the Golgi cargo binding pathway. Aim III will involve dynein-mediated virus motility. We will focus on adenovirus as a structurally simple pathogenic form of dynein cargo. We will extend and complete our efforts to identify the interacting partners between dynein and the adenovirus capsid. Together these studies have important implications for understanding many aspects of normal and abnormal cell and developmental biology. The final aim has particular significance for the control of acute viral infection and the design of gene delivery vectors and strategies.