Assembly of Cytoskeletal Proteins into the Cleavage Furrow

这个项目的重点是收缩装置 负责动物细胞的胞质分裂。 这些目标 了解肌动蛋白和肌球蛋白是如何组装成 形成卵裂沟的收缩单位，并确定 这些单位是如何附着在细胞膜上的 荧光 将注射肌动蛋白、肌球蛋白、talin和黏着斑蛋白的类似物 并将其并入卵裂沟中 分析了 肌动蛋白的组装和分解动力学， 肌球蛋白在沟将被测量和特定的抑制剂 引入来检验这两种蛋白质是 同时招募但彼此独立。 荧光肌动蛋白和肌球蛋白的短期掺入 类似物在有丝分裂将被测量，以确定是否肌动蛋白 和肌球蛋白被优先纳入赤道 皮层 talin和黏着斑蛋白的抑制剂将被引入到 有丝分裂细胞来测试这些蛋白质在有丝分裂细胞中发挥作用的想法。 将肌动蛋白丝附着在卵裂沟膜上。 肌动蛋白和肌球蛋白类似物的光活化和光漂白 将被用来确定是否有肌动蛋白的皮层流动 和肌球蛋白从两极到赤道皮层形成 卵裂沟 所提出的实验的意义在于， 描述肌动蛋白和肌球蛋白如何组装成 收缩器官是人类最重要的器官之一 生物学的基本过程：细胞分裂。

The focus of this project is on the contractile apparatus responsible for cytokinesis in animal cells. The goals are to understand how actin and myosin are assembled into the contractile units that form the cleavage furrow, and to determine how these units are attached to the cell membrane. Fluorescent analogues of actin, myosin, talin, and vinculin will be injected into living cells and their incorporation into cleavage furrows analyzed. The kinetics of assembly and disassembly of actin and myosin in the furrow will be measured and specific inhibitors introduced to test the hypothesis that the two proteins are recruited co-temporaneously but independently of one another. Short term incorporation of fluorescent actin and myosin analogues during mitosis will be measured to determine if actin and myosin are incorporated preferentially into the equatorial cortex. Inhibitors of talin and vinculin will be introduced into mitotic cells to test the idea that these proteins play a role in attaching actin filaments to the cleavage furrow membrane. Photoactivation and photobleaching of actin and myosin analogues will be used to determine if there is a cortical flow of actin and myosin from the poles to the equatorial cortex to form the cleavage furrow. The significance of the proposed experiments lies in their potential for describing how actin and myosin are assembled into the contractile apparatus that is essential for one of the most basic of biological processes: cell division.