TARGETED ASSEMBLY OF DYNEIN ON FLAGELLAR MICROTUBULES

微管和相关的分子马达负责多种 细胞运动正确实现这些目标的要求之一是 功能是微管相关分子的正确组装， 特别是机械化学发动机。例如， 细胞质动力蛋白到着丝粒有助于染色体分离 在细胞分裂期间。另一个突出的例子是， 动力蛋白必须定位于正确的位置， 调节纤毛/鞭毛运动，这对功能至关重要 输卵管和呼吸道上皮细胞中的精子。的失败 动力蛋白的精确靶向可能会转化为癌症， 不孕、呼吸功能障碍或感觉障碍。 然而，在这方面， 目前还不太清楚的是， 细胞中的正确位置。本课题主要研究 内动力蛋白臂作用于衣原体特异位点的机制 鞭毛微管我们的方法包括：1）表征 内动力蛋白臂的功能域，负责靶向 通过研究内臂动力蛋白II的再结合及其 Il亚基的表达。2)抗II抗体 中间链将被提升以识别位置和组装 伊尔的极性。3)识别参与对接位点的蛋白质， 使用化学交联剂和抗体的II结合。4)的cDNA Il的中间链将被克隆和测序，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.