MOLECULAR INTERACTIONS IN MOVING AXONEMES

移动轴丝中的分子相互作用

• 批准号: 6636010
• 负责人: GIANNI PIPERNO
• 金额: $ 31.27万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-04-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6636010
• 关键词: Chlamydomonas; actins; autoradiography; cilium /flagellum motility; confocal scanning microscopy; dynein ATPase; electron microscopy; fluorescence microscopy; immunoelectron microscopy; immunoprecipitation; intermolecular interaction; microtubules; molecular cloning; phenotype; phosphorylation; polymerase chain reaction; protein binding; protein structure function; protein transport; tissue /cell culture; video microscopy

DESCRIPTION (applicant's description): The goal of this project is to understand the mechanism of the protein transport process that is required for assembly, function and maintenance of Chlamydomonas flagella. This process is referred to as intraflagellar transport (IFT) and is conserved in motile and immotile cilia of multicellular organisms. Therefore, the information obtained on the IFT in a model system such as Chlamydomonas, likely is pertinent to the understanding of the biogenesis of cilia, flagella and sensory outer segments in humans. Dysfunctions of immotile cilia affect sensory transduction, whereas dysfunctions of motile cilia cause respiratory ailments, sterility or developmental abnormalities such as situs inversus. The flagellar apparatus of Chlamydomonas is more accessible than chemosensory neurons of C. elegans or embryonic cilia of sea urchin or mouse, which are other systems used to study the IFT. The IFT involves protein particles, referred to as IFT particles, that move continuously and bidirectionally between the basal bodies and the distal end of flagella. These IFT particles are recycled and change size and/or structure at both flagellar extremities. For these reasons we propose a functional scheme, in which the IFT cycle is divided in four phases to account for anterograde and retrograde motion of the particles and the functions served by the particles at both ends of flagella. The long-term objective of this proposal will be approached with the following four specific aims: A. To identify sets of temperature-sensitive mutants each defective in one of the four phases of the IFT cycle. B. To identify the proteins of an IFT particle that bind to cargo and/or molecular motors. C. To clone genes encoding relevant proteins of the machinery carrying out the IFT. D. To identify the proteins that use IFT particles to reach their final location within flagella. Pivotal for the development of this project is a procedure used for quantitative analysis of sequences of microscopic images that allows tracking of the motion of IFT particles for periods of several seconds in vivo. This approach is leading to the identification of distinct mutants of anterograde or retrograde IFT as well as mutants of the basal body or distal end of flagella.

描述（申请人的描述）：本项目的目标是 了解蛋白质转运过程的机制， 衣原体鞭毛的组装、功能和维持。这个过程是 称为鞭毛内运输（IFT），并在运动和 多细胞生物的不动的纤毛。因此，获得的信息 在一个模型系统，如衣原体IFT，可能是有关的 了解纤毛、鞭毛和感觉外节的生物发生 在人类身上。不动纤毛的功能障碍影响感觉传导，而 运动纤毛功能障碍导致呼吸系统疾病、不育或 发育异常，如内脏反位。 披衣菌的鞭毛器比化学感受器更易接近 C.或海胆或小鼠的胚胎纤毛， 用于研究IFT的其他系统。IFT涉及蛋白质颗粒， 被称为IFT颗粒，其连续且双向地移动 位于鞭毛的基部和末端之间。这些IFT粒子 被回收并改变鞭毛两端的大小和/或结构。 出于这些原因，我们提出了一个功能方案，其中IFT循环是 分为四个阶段，以解释顺行和逆行运动的 鞭毛两端的颗粒及其功能。 本建议的长期目标将从以下方面着手 四个具体目标：A.为了鉴定温度敏感的突变体， IFT循环的四个阶段之一出现故障。B。识别 IFT颗粒的蛋白质结合到货物和/或分子马达。C.到 克隆编码执行IFT的机器的相关蛋白质的基因。 D.为了鉴定使用IFT颗粒到达其最终目的的蛋白质， 鞭毛内的位置。 本项目的开发过程是一个用于 对显微图像序列进行定量分析， IFT粒子在体内几秒钟内的运动。这 这种方法导致了顺行或逆行的不同突变体的鉴定， 逆行IFT以及鞭毛基体或末端的突变体。