MOLECULAR BIOLOGY OF MICROTUBULE-INTERACTING PROTEINS

我们的长期目标是鉴定和鉴定 合成微管并与其相互作用。微管是人体必需的 细胞分裂、细胞结构和细胞器在细胞中的运动。 它们也是医学上重要的抗真菌和驱虫药的靶标。 药物和癌症化疗药物。尽管……很重要 微管，许多对功能起关键作用的蛋白质 微管的数量是未知的，我们的目标是确定其中的一些 蛋白质。我们的总体策略是首先通过 识别编码它们的基因，然后使用这些基因来识别 确定这些蛋白质的特性并确定它们的功能。我们有 发现了一种这样的蛋白质，伽马微管蛋白，现在我们希望测试我们的 GMMA微管蛋白作为微管核因子的假说 集合。我们将使用丝状真菌，尼杜拉曲霉来制作我们的 研究是因为它是执行分子进化的极佳有机体 我们提议进行遗传和生化实验。具体来说，我们将1) 创造新的伽马微管蛋白基因条件性致死突变等位基因 并确定这些突变对微管组装的影响 体外和体内，2)确定伽马微管蛋白抗体或突变 将阻断来自微管组织中心的微管组装和 3)纯化和鉴定γ-微管蛋白。识别额外的基因 微管功能所需的，我们建议使用 将允许我们克隆编码蛋白质的基因，这些蛋白质与 微管，是组成微管组织的蛋白质 中心和与伽马微管蛋白相关或相互作用的蛋白质。

Our long term objective is to identify and characterize proteins that compose and interact with microtubules. Microtubules are essential for cell division, cell structure, and movement of organelles through cells. They are also targets for medically important antifungal and anthelmintic agents and for cancer chemotherapy agents. In spite of the importance of microtubules, many of the proteins that are essential for the functioning of microtubules are unknown and it is our goal to identify some of these proteins. Our general strategy is to identify such proteins by first identifying the genes that encode them and then using the genes to identify and characterize the proteins and determine their functions. We have discovered one such protein, gamma tubulin, and we now wish to test our hypothesis that gmma tubulin functions as a nucleator of microtubule assembly. We will use tbe filamentous fungus, Aspergillus nidulans for our studies because it is an excellent organism for carrying out the molecular genetic and biochemical experiments we propose. Specifically we will 1) create new conditionally lethal mutant alleles of the gamma tubulin gene and determine the effects of these mutations on microtubule assembly in vitro and in vivo, 2) determine if gamma tubulin antibodies or mutations will block assembly of microtubules from microtubule organizing centers and 3) purify and characterize gamma tubulin. To identify additional genes required for microtubule functioning, we propose to use procedures that will allow us to clone genes that encode proteins that co-purify with microtubules, proteins that are components of the microtubule organizing center and proteins that are related to, or interact with, gamma tubulin.