MECHANISM AND CONTROL OF MICROTUBULE ASSEMBLY

微管组装的机制和控制

• 批准号: 3272748
• 负责人: GARY G BORISY
• 金额: $ 18万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-04-01 至 1988-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3272748
• 关键词: HeLa cells; brain; centrosome; colchicine; cytoplasm; dissection; electron microscopy; human tissue; immunoelectron microscopy; immunofluorescence technique; microinjections; mitotic spindle apparatus; neoplastic cell; polymerization; radiotracer; synchronous cell division; tissue /cell culture; tritium

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.

微管是动态结构，用于组织细胞质 相间单元格并提供结构框架，并可能 有丝分裂细胞中主轴的动力。 广泛的目标 该建议是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取决于 动态属性。 将使用各种实验方法 包括免疫荧光，显微外科手术，荧光和触觉标记 蛋白质，显微注射和荧光光漂白恢复。 这 主要目的是确定在 微管网络在相间和有丝分裂之间的过渡中；到 确定微管蛋白亚基交换的程度和地点 微管与微管相关的蛋白质：测试 微管踏板的存在和重要性；并通过使用 小管蛋白 - 钙化复合物是微管蛋白交换的特定抑制剂， 测试后期运动对微管拆卸的依赖性。 小说 包括触觉介导的免疫electorn显微镜和 荧光光漂白的荧光标记蛋白掺入 将使用微管聚合物。 结果应有所帮助 阐明微管的精确性质和细胞意义 不稳定。 结果也应与机制直接相关 调节正常细胞中的分裂。 对正常细胞的理解 分裂可以有助于分析机制的癌细胞 限制部门异常。