Regulation of Flagellar Assembly in Chlamydomonas

衣藻鞭毛组装的调控

• 批准号: 6867285
• 负责人: PAUL A. LEFEBVRE
• 金额: $ 29.17万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6867285
• 关键词: Chlamydomonas; RNA interference; affinity chromatography; antibody; biological signal transduction; cell cycle; cilium; cyclin dependent kinase; enzyme substrate; flagellum; gene interaction; genetic regulation; immunoprecipitation; mitogen activated protein kinase; molecular genetics; mutant; phosphorylation; protein protein interaction; protozoal genetics; two dimensional gel electrophoresis; yeast two hybrid system

DESCRIPTION (provided by applicant): The long-term goal of this research is to understand the mechanisms eukaryotic cells use to regulate the assembly of cilia and flagella. The flagella of Chlamydomonas are maintained at a constant length, but mutations in four different genes: LF1, LF2, LF3 and LF4, cause the cells to lose control of assembly and grow flagella up to three times normal length. Two of these genes encode members of well-studied gene families: a MAP kinase and a kinase of the CDK family. Analysis of the four LF genes leads to the conclusion that LF4, a novel MAP kinase, acts to enforce flagellar length control by shortening the flagella. LF1, LF2 and LF3 appear to act together to regulate length, perhaps by regulating retrograde intraflagellar transport (IFT). During the next project period the specific aims of this project will address the following questions:l) What proteins regulate the MAP kinase enzyme encoded by the LF4 gene, and what proteins does LF4p phosphorylate? 2) How do the proteins in the putative LF1/LF2/LF3 cytoplasmic complex interact, and how does this complex regulate the LF2 protein kinase? 3) What are the protein substrates of the LF2 protein kinase and how does phosphorylation of these targets regulate flagellar length? 4) Do the long-flagella mutants show alterations in intraflagellar transport (IFT)? Cilia and flagella have long been known to play key roles in processes that involve the movement of fluids over surfaces or the movement of cells through fluids. More recently it has become clear that cilia and flagella have critical functions early in development. Defects in the assembly of cilia and flagella have been connected to polycystic kidney disease retinal degeneration, situs inversus, and other problems in mammalian development. Understanding the regulation of flagellar assembly in Chlamydomonas provides a powerful model for understanding this assembly in mammalian systems.

描述（由申请人提供）：这项研究的长期目标是了解真核细胞用于调节纤毛和鞭毛组装的机制。衣原体的鞭毛保持恒定的长度，但四个不同基因的突变：LF1，LF2，LF3和LF4，导致细胞失去对组装的控制，并长出鞭毛长度为正常长度的三倍。这些基因中的两个编码了良好的基因家族的成员：一个地图激酶和CDK家族的激酶。对四个LF基因的分析得出的结论是，LF4是一种新型的MAP激酶，可通过缩短鞭毛来实施鞭毛长度控制。 LF1，LF2和LF3似乎通过调节逆行内部转运（IFT）来调节长度。在下一个项目期间，该项目的具体目的将解决以下问题：l）哪些蛋白质调节LF4基因编码的MAP激酶酶，以及LF4P磷酸化的哪些蛋白质？ 2）假定的LF1/LF2/LF3细胞质复合物中的蛋白质如何相互作用，该复合物如何调节LF2蛋白激酶？ 3）LF2蛋白激酶的蛋白质底物是什么？这些靶标的磷酸化如何调节鞭毛长度？ 4）长flagella突变体是否显示出flagellar内转运（IFT）的改变？早就知道纤毛和鞭毛在涉及流体在表面上运动或细胞通过液体运动的过程中起关键作用。最近，很明显，纤毛和鞭毛在开发的早期就具有关键功能。纤毛和鞭毛组装中的缺陷已与多囊性肾脏疾病视网膜变性，现场反相和哺乳动物发育中的其他问题有关。了解衣原体中鞭毛大会的调节为理解哺乳动物系统中的这一组件提供了有力的模型。