Mutational Studies of Processive Myosin Motors

描述（由申请人提供）：这是母基金GM 078097，进行性肌球蛋白马达的突变研究的竞争性修订。它试图通过提出在培养的哺乳动物细胞中进行马达蛋白的实验来扩大原始申请的范围，与纯粹涉及体外表征的母申请相反。此外，它既发挥了创造就业机会的作用（将雇用一名新的博士后助理），又通过这种人力投入加快了研究的克里思。第一个目标是确定肌球蛋白V的哪些特征对于它有效地通过构成活细胞中肌动蛋白皮质的网络是必不可少的。将在COS-7细胞中确定马达的占空比和每个货物的马达数量对于基于肌球蛋白V的货物运输的重要性。其次，我们将研究如何移交货物，从微管上的驱动蛋白为基础的运动，以肌球蛋白V为基础的运输肌动蛋白，是促进细胞内。驱动蛋白和肌球蛋白V将附着在相同的货物上，以跟随从细胞中心到外周的运输。这项工作的影响将是建立在何种程度上，我们与肌球蛋白V和驱动蛋白在体外研究的基本属性也在细胞内的操作。许多人类疾病是由于货物运输沿着肌动蛋白或微管的破坏，或者是由于疾病蛋白充当这些运输蛋白的货物。更好地理解分子马达在细胞中沿着它们的轨道运动的机制对于进一步理解这些疾病的基础至关重要。 公共卫生相关性：许多人类疾病是由于货物运输沿着肌动蛋白或微管的破坏，或者是由于疾病蛋白充当这些运输蛋白的货物。更好地理解分子马达在细胞中沿着它们的轨道运动的机制对于进一步理解这些疾病的基础至关重要。

DESCRIPTION (provided by applicant): This is a competitive revision of parent grant GM078097, Mutational Studies of Processive Myosin Motors. It seeks to expand the scope of the original application by proposing experiments with motor proteins in cultured mammalian cells, in contrast to the parent application which purely involved in vitro characterization. Moreover, it fulfills both the role of job creation (a new postdoctoral associate will be hired) as well as accelerating the tempo of research by this input of manpower. The first aim seeks to establish what features of myosin V are essential for it to effectively maneuver through the meshwork that constitutes the actin cortex in a living cell. The importance of duty cycle of the motor and motor number per cargo for myosin V based cargo-transport will be determined in COS-7 cells. Secondly, we will investigate how the handoff of cargo, from kinesin-based movement on microtubules to myosin Vbased transport on actin, is facilitated within the cell. Kinesin and myosin V will be attached to the same cargo to follow transport from the cell center out to the periphery. The impact of this work will be to establish the extent to which the basic properties we associate with myosin V and kinesin from in vitro studies are also operative within the cell. A number of human diseases result from disruption of cargo transport along actin or microtubules, or from disease proteins acting as cargo for these transport proteins. A better understanding of the mechanism by which molecular motors move along their track in the cell is critical to further understand the basis of these diseases. PUBLIC HEALTH RELEVANCE: A number of human diseases result from disruption of cargo transport along actin or microtubules, or from disease proteins acting as cargo for these transport proteins. A better understanding of the mechanism by which molecular motors move along their track in the cell is critical to further understand the basis of these diseases.