Actin Myosin Interactions in Cell Motility

描述（由申请人提供）：我的长期目标是了解胞质分裂的机制。我们将追求互补的细胞，生物化学和遗传策略，使用裂殖酵母裂殖酵母，以解决以下具体目标：项目1。收缩环蛋白的动力学：这些活细胞中分子的定量实验将提供破译收缩环组装途径所需的参数值。我们将测量胞质分裂蛋白质之间的细胞质和收缩环交换的速率，确定肌动蛋白丝是否影响每个收缩环蛋白的动态，并确定胞质分裂蛋白质是否影响彼此的动态。项目2.胞质分裂蛋白在前体节点宽带中的组装：这些实验将阐明组装收缩环前体的生化途径。我们将确定结合皮质和下游蛋白质的苯胺Mid1p的结构域，以及胞质分裂蛋白如何与Mid1p相互作用。项目3。前体节点转化为收缩环：这项工作将提供理解收缩环组装第二步所需的信息。我们将研究Cdc12p如何控制从节点的肌动蛋白丝的组装，表征机械过程，转换，宽带节点成收缩环，表征DCS蛋白Rng3p和磷酸化如何影响Myo2丝的结构，并确定胞质分裂蛋白是如何组织在宽带和收缩环。项目4。收缩环的收缩和分解：这些实验将帮助我们了解细胞如何触发收缩环的收缩，以及收缩时环如何分解。我们将研究SIN通路如何触发收缩环的收缩，探索限制收缩环收缩速率的因素，并确定收缩环在收缩过程中如何分解。

DESCRIPTION (provided by applicant): My long range goal is to understand the mechanism of cytokinesis. We will pursue complementary cellular, biochemical and genetic strategies using the fission yeast Schizosaccharomyces pombe to address the following specific aims: Project 1. Dynamics of contractile ring proteins: these quantitative experiments on molecules in live cells-will provide the parameter values required to decipher the pathway of contractile ring assembly. We will measure the rates that cytokinesis proteins exchange between the cytoplasm and contractile rings, determine if actin filaments influence the dynamics of each contractile ring protein and determine if cytokinesis proteins influence each other's dynamics. Project 2. Assembly of cytokinesis proteins in the broad band of precursor nodes: these experiments will clarify the biochemical pathway that assembles the precursor of the contractile ring. We will determine the domains of anillin Mid1p that bind the cortex and downstream proteins and how cytokinesis proteins interact with Mid1p and with each other. Project 3. Transformation of precursor nodes into a contractile ring: this work will provide information required to understand the second step in the assembly of the contractile ring. We will investigate how Cdc12p controls the assembly of actin filaments from nodes, characterize the mechanical process that converts, a broad band of nodes into a contractile ring, characterize how DCS protein Rng3p and phosphorylation affect the structure of Myo2 filaments and determine how cytokinesis proteins are organized in the broad band and the contractile ring. Project 4. Constriction and disassembly of the contractile ring: these experiments will help us understand how cells trigger constriction of the contractile ring and how the ring disassembles as it constricts. We will investigate how the SIN pathway triggers constriction of the contractile ring, explore the factors that limit the rate of constriction of contractile rings and determine how the contractile ring disassembles during constriction.