Intermediate Filaments: Motility, Motors and Cytoskeletal Interactions

项目摘要-项目2（Gelfand） 这个提议的目的是研究波形蛋白中间丝与另外两个中间丝之间的动态相互作用。 细胞骨架，微管和肌动蛋白微丝的组成部分，以及VIF与 膜细胞器我们将研究中间丝如何沿沿着微管移动， 微管发动机来驱动这种运动。我们将确定波形蛋白相互作用的生化基础 与微管马达，并确定参与这些相互作用的调节的分子机制， 例如波形蛋白的翻译后修饰。我们还将发现波形蛋白的分子机制 与肌动蛋白微丝和它们的束的相互作用以及与粘着斑的组分的相互作用。我们将测试如果 VIF与粘着斑的相互作用影响细胞-基质相互作用的稳定性以及VIF粘着斑中波形蛋白的作用 粘附调节细胞运动性。我们将确定肌动蛋白和中间丝之间的分子连接 并研究这种关联的功能含义。我们还将发现中间纤维是如何侵入病灶的 粘附，粘着斑蛋白将它们连接到位，以及中间丝如何调节粘附 周转最后，波形蛋白丝在膜细胞器运输中的作用将通过使用 两个模型系统，线粒体和色素细胞器（黑素体）在黑色素细胞，因为这两个 这些细胞器含有结合的PKA，PKA依赖性磷酸化可能与 中间丝动力学和细胞器运输。

Project Summary—Project 2 (Gelfand) The goal of this proposal is to study dynamic interactions of vimentin intermediate filaments with two other components of the cytoskeleton, microtubules and actin microfilaments, as well as interactions of VIF with membrane organelles. We will study how intermediate filaments move along microtubules and identify microtubule motors that power this movement. We will determine biochemical basis of vimentin interactions with microtubule motors and identify molecular mechanisms involved in the regulation of these interactions, such as posttranslational modifications of vimentin. We will also find molecular mechanisms of vimentin interaction with actin microfilaments and their bundles and with components of focal adhesions. We will test if interactions of VIF with focal adhesions affect stability of cell-substrate interactions and how vimentin in focal adhesions modulates cell motility. We will identify molecular linkers between actin and intermediate filaments and study functional implications of this association. We will also find how intermediate filaments invade focal adhesions, which focal adhesion proteins link them in place and how intermediate filaments regulate adhesion turnover. Finally, the role of vimentin filaments in transport of membrane organelles will be investigated using two model systems, mitochondria and pigment organelles (melanosomes) in melanophores, because both of these organelles contain bound PKA and PKA-dependent phosphorylation is likely regulated in concert with intermediate filament dynamics and organelle transport.