INTRAFLAGELLAR TRANSPORT IN CHLAMYDOMONAS REINHARDTII

莱因衣藻的鞭内运输

基本信息

批准号：
7585649
负责人：
DOUGLAS GENE COLE
金额：
$ 26.99万
依托单位：
UNIVERSITY OF IDAHO
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-08-01 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7585649
关键词：
Address Affect Animal Model Anosmia Architecture Back Biochemical Biological Biological Assay Caenorhabditis elegans Carrier Proteins Cell surface Cells Chemicals Chlamydomonas Chlamydomonas reinhardtii Cilia Complement Complex Defect Disease Distal Docking Dynein ATPase Electron Microscopy Environment Enzymes Female infertility Flagella Fluorescence Funding Genes Genetic Genetic Transcription Goals Green Algae Housing Hydrocephalus Immunoprecipitation Insertional Mutagenesis Kinesin Length Link Liquid substance Maintenance Microarray Analysis Molecular Molecular Motors Motor Movement Mucous body substance Nematoda Odorant Receptors Organelles Orthologous Gene Partner in relationship Phosphoenolpyruvate Carboxylase Photoreceptors Polycystic Kidney Diseases Primary Ciliary Dyskinesias Proteins Proteomics Protozoa RNA Interference Radial Relative (related person)Research Research Personnel Retinal Degeneration Role Sensory Site Source Stimulus Structure Swimming Syndrome Temperature Testing Trachea Tubulin Upper arm Visible Radiation Yeasts appendage base cell motility cell type comparative crosslink design human disease male man mutant neuronal cell body particle polypeptide programs sperm cell transcription factor yeast two hybrid system

项目摘要

Eukaryotic cilia and flagella are ancient cellular appendages that have been adapted for motile and sensory functions. Motile forms of these organelles are capable of propelling some cells like sperm and protozoa through a liquid environment while other cells like the ciliated trachea of man use the coordinated beating of many cilia to propel a liquid or mucous environment over the surface of the cells. Nonmotile cilia have been adapted to sense a wide range of stimuli. Classic examples include the photoreceptors which are highly modified cilia that can sense visible light and the olfactory cilia which are highly enriched in odorant receptors. Because of their important roles in both motility and sensory transduction, defects in cilia and flagella have been intimately linked with a number of human diseases including retinal degeneration, immotilie cilia and Kartagener's syndromes, male and female infertility, hydrocephalus and anosmia, Bardet- Beidl syndrome and one of the most commongenetic diseases in man, polycystic kidney disease. Focusing on how cells build these organelles, we study intraflagellar transport (IFT) which is required for the assembly and maintenance of these structures. IFT is characterized by the movement of protein particles along the long axis of the organelle, both out to the tip (anterograde IFT driven by kinesin-2) and back to the cell body (retrograde IFT driven by cytoplasmic dynein 1b/2). A primary function of IFT is to transport axonemal building blocks out to the distal tip which serves as the site of assembly for the organelle. The model organism for the study of IFT is the unicellular biflagellate green alga, Chlamydomonas reinhardtii. Biochemical analysis of the 17 proteins found in the Chlamydomonas IFT particles has begun to reveal their complex oligomeric organization. Continuing on our previous study of the IFT particles, our specific aims are: (1) to further characterize the architecture of IFT particles; to identify how the 17 proteins assemble into complexes; (2) to characterize the interaction of the IFT particles with the IFT motor proteins, kinesin-2 and cytoplasmic dynein 1b; (3) to directly visualize the transport of hypothesized IFT cargo including tubulin and radial spoke complexes; and (4) to address the function of separate IFT particle proteins. An important aspect of Aim 4 will involve the creation of 900 or more motility mutants; currently we lack mutants in more than 1/2 of the IFT particle proteins.

真核生物的纤毛和鞭毛是古老的细胞附属物，已适应运动和运动的需要。感觉功能。这些细胞器的运动形式能够推动一些细胞，如精子和原生动物通过液体环境，而其他细胞（例如人类的纤毛气管）则使用协调的许多纤毛的跳动以推动细胞表面上的液体或粘液环境。不动的纤毛已经适应了感知广泛的刺激。经典的例子包括光感受器高度修饰的纤毛可以感知可见光，嗅觉纤毛富含气味受体。由于纤毛和感觉转导中的重要作用，纤毛和鞭毛与许多人类疾病密切相关，包括视网膜变性、纤毛不动和卡塔格纳综合征、男性和女性不育症、脑积水和嗅觉丧失、Bardet- 贝德尔综合征和人类最常见的遗传性疾病之一——多囊肾。我们专注于细胞如何构建这些细胞器，研究所需的鞭毛内运输（IFT）用于这些结构的组装和维护。 IFT的特点是蛋白质的运动颗粒沿着细胞器的长轴，均到达尖端（由驱动蛋白-2 驱动的顺行 IFT）和回到细胞体（由细胞质动力蛋白 1b/2 驱动的逆行 IFT）。 IFT 的主要功能是将轴丝构件运输到远端，作为细胞器的组装部位。 IFT 研究的模式生物是单细胞双鞭毛绿藻 Chlamydomonas 莱茵哈蒂。衣藻 IFT 颗粒中发现的 17 种蛋白质的生化分析已开始揭示其复杂的寡聚组织。继续我们之前对 IFT 粒子的研究，我们具体目标是：（1）进一步表征IFT粒子的结构；确定 17 种蛋白质如何组装成复合体； (2) 表征 IFT 颗粒与 IFT 运动蛋白的相互作用，驱动蛋白-2 和细胞质动力蛋白 1b； (3) 直接可视化假设的 IFT 货物的运输包括微管蛋白和放射状辐条复合物； (4) 解决单独 IFT 粒子的功能蛋白质。目标 4 的一个重要方面将涉及创建 900 个或更多的运动突变体；目前我们超过 1/2 的 IFT 颗粒蛋白缺乏突变体。