TARGETED ASSEMBLY OF DYNEIN ON FLAGELLAR MICROTUBULES

微管和相关的分子马达负责 细胞运动。要正确实现这些目标，其中一个要求 功能是微管相关分子的正确组装， 尤其是机械力化学马达。例如，绑定 细胞质动力蛋白对着丝点的作用有助于染色体分离 在细胞分裂过程中。另一个引人注目的例子是4种亚型 动力蛋白必须定位在正确的位置才能达到高度 调节纤毛/鞭毛运动，这对功能至关重要 输卵管和呼吸道的精子和上皮组织。失败的原因是 动力蛋白的精确靶向可能会转化为癌症， 不孕不育、呼吸功能障碍或感觉障碍。然而， 关于将马达引导到 单元格中的正确位置。本课题研究的是目标定位问题 衣藻体内动力蛋白臂作用于特定部位的机制 鞭毛微管。我们的方法包括：1)表征 内动力蛋白臂的功能域，Il负责靶向 研究内臂动力蛋白Il及其受体的重新结合组装 在体外缺少Il的轴丝亚基。2)抗II抗体 中间链将被提升以确定位置和组装 伊利诺伊的极性。3)确定与对接位置有关的蛋白质 使用化学交联剂和抗体结合。4)该基因的编码区序列 将克隆和测序Il中间链，以便于 进一步研究靶向组装的分子机制。

Microtubules and associated molecular motors are responsible for multiple cellular motion. One of the requirements to properly achieve these functions is correct assembly of microtubule-associated molecules, particularly the mechanochemical motors. For example, the binding of cytoplasmic dynein to kinetochores contribute to chromosome segregation during cell division. The other striking example is that 4 isoforms of dynein must be targeted to the correct location to achieve highly regulated ciliary/flagellar motility, which is critical for the function of sperm and epithelium in oviduct and respiratory tract. The failure of precise targeting of dynein could potentially be translated into cancer, infertility, respiratory dysfunction or sensory impairment. However, little is yet known about the molecular mechanism which direct motors to the correct position in the cell. This project is to study the targeting mechanism of inner dynein arms to specific sites on Chlamydomonas flagellar microtubules. Our approaches include: 1) Characterize the functional domain of inner dynein arm, Il responsible for targeted assembly by studying the rebinding of inner arm dynein Il and its subunits to axoneme lacking Il in vitro. 2) Antibodies against II intermediate chains will be raised to identify the location and assembly polarity of Il. 3) Identify the proteins involved in the docking site for II binding using chemical cross linker and antibodies. 4) The cDNA of the intermediate chain of Il will be cloned and sequenced to facilitate further study the molecular mechanism of targeted assembly.