Cell Polarity and Spindle Position

项目描述（申请人提供）：该项目的目标是全面了解出芽酵母sacaromyces cerevisiae中导致有丝分裂纺锤体定位的分子机制。在许多细胞类型中，这一过程是不对称细胞分裂和发育的基础。纺锤体定位涉及星状微管（MTs）与细胞皮层极化位点的相互作用。在许多情况下，这个过程也需要肌动蛋白细胞骨架。MT在细胞皮层的捕获与在着丝点的捕获有相似之处，一些蛋白质显然参与了这两个过程。因此，了解纺锤体定位的机制也可能影响基因组稳定性的维持。因此，我们的研究通过对发展和癌症的影响来解决影响人类健康的机制。我们的方法利用遗传、生化和成像实验。它还受益于我们最近解决的系统的一个组成部分的结构，Bnil FH2结构域。在我们之前的研究工作的基础上，表征了与mt的正端相关的蛋白质和相关的蛋白质，以及纺锤体定向所需的肌动蛋白组装方面，我们提出了以下目标的实验：目的1：如何产生定向前期纺锤体的力？目标2：动力蛋白是如何靶向作用部位的？动力蛋白的活性是如何调控的？目的3：体内形成蛋白依赖的肌动蛋白组装是如何调节的？

DESCRIPTION (provided by applicant): The goal of this project is to comprehensively understand the molecular mechanisms leading to the positioning of the mitotic spindle in the budding yeast Saccaromyces cerevisiae. This process underlies asymmetric cell division and development in many cell types. Spindle positioning involves the interaction of astral microtubules (MTs) with polarized sites on the cell cortex. In many cases, this process also requires the actin cytoskeleton. The capture of MT at the cell cortex shares similarities with the capture of MTs at the kinetochore and some proteins are clearly involved in both processes. Thus, understanding mechanisms for spindle positioning may also impact upon the maintenance of genome stability. Our studies therefore address mechanisms that impact on human health through implications for development and cancer. Our approach utilizes genetic, biochemical, and imaging experiments. It also benefits from our recently solved structure of one component of the system, the Bnil FH2 domain. Building on our previous work to characterize proteins associated with the plus ends of MTs and proteins involved and aspects of actin assembly that are required for spindle orientation we propose experiments with the following aims: Aim1: How is the force generated to orient preanaphase spindles? Aim 2: How is dynein targeted to its site of action and how is dynein activity regulated? Aim 3: How is formin-dependent actin assembly regulated in vivo?