MOLECULAR ANALYSIS OF CYTOSKELETAL INTERACTIONS

这项拟议的研究的目标是了解细胞骨架是如何 针对不同的职能进行组织，以及该组织以什么方式 在癌细胞中被破坏。特别注意结构 和膜细胞骨架连接的功能。拟议的研究 利用几种动物细胞模型进行分化和 致癌转化，以确定可能是关键的分子 变化发生在差异化时，这也可能是 转化基因。这些实验的一个主要焦点是分析 Ezrin相关蛋白的功能。我们已经证明了这种蛋白质 具有微管和微丝相关的特性 蛋白。例如，它以类似的方式定位到生长锥，但 与F-肌动蛋白不完全相同。但它在年增长锥体中的位置 转折取决于完整的微管。这些结果和其他结果表明 这种蛋白质可能与这两种细胞骨架相互作用 元素。Ezrin本身是与致癌相关的激酶的底物 转型和增长因素。此外，还测定了Ezrin的DNA序列 提示它具有膜结合结构域。这些加在一起， 研究结果表明，Ezrin可能在调节 细胞骨架之间的连接和质膜连接。 一系列结果认为，膜-细胞骨架的连接是 在致癌转化过程中被修饰，并在 分化细胞形态的表达及其可能介导的丢失 在变形时发生的这种形态。建议的部分 研究旨在分析这些相互作用及其相关性 对转型和分化，进行了详细的阐述。在相关工作中，其他 实验的目的是识别那些必不可少的分子。 用于表达分化的细胞骨架组织，并 为它们的功能设计和应用严格的测试。

The goal of the proposed research is to understand how the cytoskeleton is organized for differentiated function, and in what ways that organization is disrupted in cancer cells. Particular attention is paid to the structure and function of the membranecytoskeleton connection. The proposed research takes advantage of several animal cell models for differentiation and oncogenic transformation to identify molecules that may be crucial for the changes occur upon differentiation and which may also be the targets of transforming genes. A major focus of these experiments is an analysis of the functions of an ezrin-related protein. We have shown that this protein has properties of both a microtubule and a microfilament-associated protein. For example, it localizes to growth cones in a manner similar but not identical to that of F-actin. But its position at the growth cone in turn depends upon intact microtubules. These results and others suggest that this protein may interact with both of these two cytoskeletal elements. Ezrin itself is a substrate for kinases associated with oncogenic transformation and growth factors. In addition, the DNA sequence of ezrin suggests that it has a membrane binding domain. Taken together, these findings suggest that ezrin may play a crucial role in mediating the connections between the cytoskeleton and the plasma membrane connection. A range of results argues that the membrane-cytoskeleton connection is modified upon oncogenic transformation, and play an important role in the expression of differentiated cell morphology and which may mediate the loss of that morphology that occurs upon transformation. Part of the proposed research is designed to analyze these interactions, and their dependence upon transformation and differentiation, in detail. In related work, other experiments are designed to identify those molecules which are essential for the expression of differentiated cytoskeletal organization, and to design and apply rigorous tests for their function.