REGULATION OF ASYMMETRIC FLAGELLAR WAVEFORMS

不对称鞭毛波形的调节

• 批准号: 2459556
• 负责人: PAUL A. LEFEBVRE
• 金额: $ 17.21万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 1998-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2459556
• 关键词: Chlamydomonas; chemical binding; chemical models; cilium /flagellum motility; epitope mapping; gene complementation; immunoelectron microscopy; immunofluorescence technique; microtubule associated protein; molecular cloning; nucleic acid hybridization; nucleic acid sequence; phase contrast microscopy; phenotype; phosphorylation; polymerase chain reaction; protein sequence; protein structure function; restriction mapping; site directed mutagenesis; southern blotting; transmission electron microscopy

The unicellular green alga Chlamydomonas can swim with two very different flagellar waveforms. Forward swimming is accomplished using an asymmetric beat, resembling the beat of cilia. Reverse swimming is accomplished by a symmetrical waveform, similar to classic flagellar beating. The transition between the two forms is controlled by the contraction of Ca++ around the axoneme. Two different classes of mutants in Chlamydomonas have been isolated which have lost the ability to generate the asymmetric waveform: central pair apparatus defective mutants and move backward only mutants. The recent development of insertional mutagenesis in this organism has made it possible to rapidly generate mutants in many genes regulating flagellar function and to clone the affected genes. This technique will be used in this project to begin a comprehensive molecular, biochemical and ultrastructural analysis of the regulation of flagellar waveforms. The specific aims are: 1) to use insertional mutagenesis to identify and clone genes whose products are required for generation of an asymmetric waveform in Chlamydomonas flagella; 2) to identify the gene products of each cad and mbo gene by sequencing genomic and cDNA clones; 3) to localize each gene product within the flagellum; 4) to address the function of each gene product by detailed characterization of mutant phenotypes. Chlamydomonas is the primary genetic system available for the study of flagellar structure and function. Dozens of mutants affecting flagellar function have been well- characterized, and many of the genes coding for components of the microtubules, radial spoke assembly and dynein arms have been isolated. The central apparatus is perhaps the least understood region of the axoneme. This product will identify and characterize many of the proteins involved in the generation of the asymmetric flagellar waveform. This research will advance our understanding of human biology in several important ways, given the important role of cilia in respiratory function and the role of flagella in male fertility.

单细胞的绿色衣原体可以和两种完全不同的 鞭毛波形 向前游泳是用一个 不对称的搏动，类似纤毛的搏动。 反向游泳是 由对称的波形完成，类似于经典的鞭毛 心要让 两种形式之间的转换由 轴丝周围Ca++的收缩。 两类不同的突变体 在衣原体中已经被分离出来， 产生不对称波形：中心对装置故障 突变体和向后移动只有突变体。 的最新发展 在这种生物体中的插入突变使其有可能快速地 在许多调控鞭毛功能的基因中产生突变体， 受影响的基因。 这项技术将用于本项目的开始 一个全面的分子，生化和超微结构分析， 鞭毛波形的调节。 具体目标是：（1）使用 插入诱变以鉴定和克隆其产物 在衣原体中产生不对称波形所需的 鞭毛; 2）鉴定每个CAD和MBO基因的基因产物， 测序基因组和cDNA克隆; 3）定位每个基因产物 在鞭毛内; 4）通过以下方式解决每个基因产物的功能： 突变体表型的详细表征。 衣原体是 初级遗传系统可用于鞭毛结构的研究， 功能 几十个影响鞭毛功能的突变体已经很好地- 特征，和许多基因编码的组成部分， 微管，径向辐条组件和动力蛋白臂已被分离。 中央装置可能是最不了解的区域， 轴丝 该产品将识别和表征许多 参与产生不对称鞭毛波形的蛋白质。 这项研究将在几个方面促进我们对人类生物学的理解 重要的方式，鉴于纤毛在呼吸功能中的重要作用， 以及鞭毛在男性生育力中的作用。