MOLECULAR ANALYSIS OF CYTOSKELETAL INTERACTIONS

这项研究的目的是了解细胞骨架是如何 为不同的功能而组织，以及组织以何种方式 在癌细胞中被破坏。特别注意结构 以及膜与骨骼连接的功能。拟议研究 利用几种动物细胞模型进行分化， 致癌转化，以确定分子，可能是至关重要的， 在分化时发生变化，也可能是 转化基因这些实验的一个主要重点是分析 一种与埃兹蛋白相关的蛋白的功能。我们已经证明这种蛋白质 具有微管和与微管相关的 蛋白例如，它以类似的方式定位于生长锥， 与F-肌动蛋白不同。但它在生长锥中的位置 转依赖于完整的微管。这些结果和其他结果表明， 这种蛋白质可以与这两种细胞骨架相互作用， 元素埃兹蛋白本身是与致癌相关的激酶的底物。 转化和生长因子。此外，Ezrin的DNA序列 表明它有一个膜结合域。综上所述各项 研究结果表明，埃兹蛋白可能在介导 细胞骨架和质膜之间的连接。 一系列的结果表明，膜-细胞骨架连接是 在致癌转化后修饰，并在肿瘤发生中发挥重要作用。 表达分化的细胞形态和可能介导的损失 在转化过程中所发生的形态变化。部分拟议 研究旨在分析这些相互作用及其依赖性 在转化和分化的过程中，在相关工作中，其他 实验的目的是确定那些分子， 用于表达分化的细胞骨架组织，以及 设计并应用严格的功能测试。

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.