Molecular and Genetic Analysis of Flagellar Dyneins

描述（由申请人提供）：已知真核生物纤毛和鞭毛的运动性依赖于沿沿着双线微管运动的动力蛋白ATP酶马达。鞭毛动力蛋白必须被调节以在正常弯曲形成和传播期间协调它们的活性，并且另外调节拍频和波形以响应变化的生理要求。外排动力蛋白组装所必需的两个基因，ODA 7和ODA 8，最近被鉴定为LC 1的同源物，其结合HC γ动力蛋白催化结构域并可作为调节轻链。将测试Oda7p和Oda8p作为HC α和HC β调节轻链的假设功能。支架蛋白被假设为沿着沿着每个双微管的多个动力蛋白同种型之间的连接。两个候选支架蛋白基因（PF13和ODA16）已被克隆的基础上插入突变，破坏动力蛋白的组装和功能。实验中提出了确定的作用，每个基因产物在对接动力蛋白调控蛋白的双重微管表面。驱动蛋白代表了在鞭毛中发现的另一个微管相关运动蛋白家族，但它们在鞭毛运动中的作用尚不清楚。鞭毛运动的全球调节涉及双联体相关动力蛋白和中央对微管相关复合物之间的径向辐条传输的信号。两个中心对投影在这一过程中的作用将研究使用突变，破坏一个投影（cpcl）和RNAi介导的敲除在另一个投影（Klp1）中发现的驱动蛋白。中心对驱动蛋白的工作将扩展到包括所有被确定为中心对蛋白的驱动蛋白，敲低菌株将用于筛选中心对驱动蛋白基因的无效突变。

DESCRIPTION (provided by applicant): The motility of eukaryotic cilia and flagella is known to depend on dynein ATPase motors that move along doublet microtubules. Flagellar dyneins must be regulated to coordinate their activity during normal bend formation and propagation, and additionally to modulate beat frequency and waveform in response to changing physiological requirements. Two genes essential for outer row dynein assembly, ODA7 and ODA8, have recently been identified as homologs of LC1, which binds to the HC gamma dynein catalytic domain and may act as a regulatory light chain. Hypothesized functions of Oda7p and Oda8p as HC alpha and HC beta regulatory light chains will be tested. Scaffold proteins have been hypothesized to function as links between multiple dynein isoforms along each doublet microtubule. Two candidate scaffold protein genes (PF13 and ODA16) have been cloned based on insertional mutations that disrupt dynein assembly and function. Experiments are proposed to define the role of each gene product in docking dyneins to regulatory proteins on the doublet microtubule surface. Kinesins represent another family of microtubule associated motor proteins found in flagella, but their role in flagellar motility is not known. Global regulation of flagellar motility involves signals transmitted by radial spokes between doublet-associated dyneins and the central pair microtubule-associated complex. The role of two central pair projections in this process will be studied using a mutation that disrupts one projection (cpcl) and RNAi-mediated knockdown of a kinesin found in another projection (Klp1). Work on central pair kinesins will be extended to include all kinesins identified as central pair proteins, and knockdown strains will be used to screen for null mutations in central pair kinesin genes.