MOLECULAR INTERACTIONS IN MOVING AXONEMES

移动轴丝中的分子相互作用

• 批准号: 3303586
• 负责人: GIANNI PIPERNO
• 金额: $ 20.76万
• 依托单位: MOUNT SINAI SCHOOL OF MEDICINE OF CUNY
• 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 1995-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3303586
• 关键词: Chlamydomonas; actins; adenosine triphosphate; calcium binding protein; chemical binding; chemical structure; chimeric proteins; cilium /flagellum motility; complementary DNA; dynein ATPase; electron microscopy; genetic library; genome; hybridomas; immunoelectron microscopy; intermolecular interaction; laboratory mouse; microtubules; molecular cloning; nucleic acid sequence; phosphorylation; polymerase chain reaction; posttranslational modifications; protein sequence; protein structure; radionuclides; tissue /cell culture

Cilia and flagella are present in a wide variety of eukaryotic cells. Their disfunction is correlated with respiratory ailments or male sterility. This proposal has the following two long-term objectives: analyze the molecular and structural features of those axonemal components that are essential for the generation of axonemal movement and second, identify the molecular mechanisms of regulation that lead to the formation of specific bending patterns of cilia and flagella. Both objectives will be approached with the analysis of the inner dynein arms, because in vivo these structures are necessary and sufficient to generate both ciliary and flagellar types of movement in the absence of outer dynein arms. The organism used is the unicellular green alga Chlamydomonas reinhardtii. This organism allows for the analysis of the axoneme by a combination of approaches including genetics, electron microscopy and biochemical procedures. The comparison of Chlamydomonas wild-type axonemes with mutant axonemes lacking the outer dynein arms or part of the inner arms forms the basis of many experimental procedures outlined in this proposal. The comparison of inner dynein arms prepared from motile flagella with those from nonmotile flagella and analysis of defective inner arms in mutant axonemes, will be adopted to explain the function of the posttranslational modification of inner arm heavy chains. On this basis the long-term objective will be approached with the following specific aims: 1. complete the characterization of the structure and molecular composition of all forms of inner dynein arms present in Chlamydomonas axonemes; 2. develop a detailed analysis of the structures and the modifications of inner arm heavy chains and determine whether the modification of heavy chains is part of mechanism controlling axonemal motility; 3. characterize the axonemal components whose function is correlated with the restoration of flagellar activity of radial spoke-defective mutants; 4. determine whether axonemal actin and caltractin directly are involved in those changes of inner dynein arm activity that occur when Chlamydomonas axonemes pass from the ciliary to the flagellar type of motion.

纤毛和鞭毛存在于各种真核细胞中。