MECHANISM AND CONTROL OF MICROTUBULE ASSEMBLY

微管是用来组织细胞质的动态结构。 间期细胞并提供结构框架和可能的 有丝分裂细胞中纺锤体的动力。的广泛目标 这项建议是为了了解体内和体外的微管动力学。 并确定选定的有丝分裂事件是否依赖于 动态属性。我们将使用各种实验方法。 包括免疫荧光、显微手术、荧光和半抗原标记 蛋白质的微量注射和荧光光漂白回收。这个 主要目标将是确定在 间期与有丝分裂过渡阶段的微管网络 确定微管蛋白亚基交换的程度和位置 微管相关蛋白与微管：测试 微管跑步机的存在和意义；并通过使用 微管蛋白交换的特异性抑制剂微管蛋白-秋水仙碱复合体的作用 检测后期运动对微管解体的依赖性。小说 方法包括半抗原介导的免疫电子显微镜和 荧光标记蛋白质的荧光光漂白 进入微管聚合物将被使用。结果应该会有所帮助 阐明微管的确切性质和细胞意义 不稳定。这些结果还应与机制直接相关。 调节正常细胞的分裂。对正常细胞的认识 分裂可能有助于分析癌细胞的机制 限制分裂是反常的。

Microtubules are dynamic structures that serve to organize the cytoplasm in interphase cells and to provide the structural framework and possibly the motive force for the spindle in mitotic cells. The broad objectives of this proposal are to understand microtubule dynamics in vivo and in vitro and to determine whether selected mitotic events are dependent upon the dynamic properties. A variety of experimental approaches will be used including immunofluorescence, microsurgery, fluorescent and hapten tagging of proteins, microinjection and fluorescence photobleaching recovery. The principal aims will be to identify the transformations occurring in the microtubule network at the transition between interphase and mitosis; to determine the extent and sites of exchange of tubulin subunits and microtubule associated proteins with the microtubules: to test for the existence and significance of microtubule treadmilling; and through the use of tubulin-colchicine complex, a specific inhibitor of tubulin exchange, to test the dependence of anaphase motion on microtubule disassembly. Novel methods including hapten-mediated immunoelectorn microscopy and fluorescence photobleaching of fluorescently-tagged proteins incorporated into microtubule polymer will be employed. The results should help elucidate the precise nature and cellular significance of microtubule lability. The results should also be of direct relevance for mechanisms regulating division in normal cells. An understanding of normal cell division may assist in the analysis of cancer cells where the mechanisms restricting division are aberrant.