Tools for mapping the tubulin landscape

微管蛋白图谱绘制工具 项目摘要： 做 超过 信息 这 微管蛋白 微管蛋白 神经元 重组 研究 是否 新 对神经元的结构复杂性和信号传导效率至关重要。但我们 没有完全理解神经元如何在正确的地点和正确的时间建立专门的微管网络。 在过去的15年里，我们已经认识到微管内的微管蛋白亚基携带着 以翻译后修饰的形式调节网络的组装和功能。 一项提案将这一概念扩展到微管蛋白同种型的水平-编码α和β的基因家族 我们的目标是开发新的工具，使有史以来第一次可视化和操纵一个 活细胞中的同种型。 该提案将创建工具，促进我们对β-微管蛋白同种型TUBB 3的理解， 形态发生，但也将在许多实验室产生广泛的影响。我们的方法将1）创建一个 TUBB 3结合抗体Tuj 1的形式，可以与其他实验室共享， 成本，2）创建第一个用于研究活细胞中微管蛋白同种型的胞内抗体，3）确定 TUBB 3表现出偏向富集到具有PTM的神经元区域或微管，4）产生一个具有PTMs的神经元区域。 快速和选择性去除细胞中TUBB 3的工具 微管 - 这 C .这项提案的成功将为 我们未来的R 01提案将阐明调节TUBB 3定位和功能的机制 以及这些在疾病中是如何被破坏的。

Tools for mapping the tubulin landscape Project Summary: do Over information This tubulins. tubulin neuronal recombinant research whether new are essential to the architectural complexity and signaling efficiency of neurons. However, we not fully understand how neurons build specialized microtubule networks at the right place and right time. the past 15 years, we have come to appreciate that the tubulin subunits within microtubules carry in the form of posttranslational modifications that regulate the assembly and function of networks. proposal extends this concept to the level of the tubulin isotypes – the gene families that encode α - and β Our goal is to develop new tools that will enable the first-ever visualization and manipulation of a isotype in living cells. proposal will create tools that will advance our understanding of the β-tubulin isotype, TUBB3, during morphogenesis, but will also have broad impacts across many labs. Our approach will 1) create a form of the TUBB3-binding antibody Tuj1 that can be shared with other labs and bring down costs, 2) create the f irst-ever intrabody for studying a tubulin isotype in living ells, 3) determine TUBB3 exhibits biased enrichment to regions of the neuron or to microtubules with PTMs, 4) create a tool for rapid and selective depletion of TUBB3 in cells Microtubules - This c . The success of this proposal will set the stage for our future R01 proposal, which will elucidate the mechanisms that regulate TUBB3 localization and function during development, and how these are disrupted in disease.