Analysis of Intraflagellar Transport

鞭毛内转运(IFT)是真核生物鞭毛和纤毛的组装和维持所必需的，也是几乎所有真核细胞的基本特征。IFT的特征是以微管为基础的运动蛋白Kinesin II将大的蛋白质复合体从基底移动到鞭毛末端，并通过细胞质dynein 1b从鞭毛末端返回到基底。IFT颗粒不是膜结合的，这些颗粒与马达蛋白和货物相互作用的机制尚不清楚。为了开始了解这些相互作用，IFT将在体外使用纯化的马达蛋白和Cargo多肽进行重组。相关实验将集中在马达蛋白与货运多肽的结合以及这些多组分IFT复合体的组装。使用参与IFT的逆行马达的温度敏感突变体的其他实验将重点放在IFT作为负控制鞭毛基因表达的信号转导系统的一个组件上。PI将把研究项目与细胞生物学的本科课程结合起来。此外，该项目还将为本科生和研究生提供研究培训。

Intraflagellar transport (IFT) is essential for the assembly and maintenance of eukaryotic flagella and cilia and is a fundamental characteristic of almost all eukaryotic cells. IFT is characterized by the movement of large protein complexes from the basal body to the flagellar tip by the microtubule-based motor protein kinesin II and from the tip back to the basal body by cytoplasmic dynein 1b. The IFT particles are not membrane bound, and the mechanism by which these particles interact with motor proteins and cargo are unknown. In order to begin understanding these interactions, IFT will be reconstituted in vitro using purified motor proteins and cargo polypeptides. Related experiments will focus on the binding of motor proteins to cargo polypeptides and the assembly of these multi-component IFT complexes. Additional experiments using temperature-sensitive mutants for the retrograde motor involved in IFT will focus on IFT as a component of a signal transduction system that negatively controls expression of flagellar genes. The PI will integrate the research project with undergraduate courses in cell biology. Moreover, the project will provide research training for undergraduate and graduate students.