Biochemistry and Function of C. elegans RNA helicase A

线虫 RNA 解旋酶 A 的生物化学和功能

• 批准号: 6897391
• 负责人: KATHERINE M WALSTROM
• 金额: $ 21.75万
• 依托单位: NEW COLLEGE OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6897391
• 关键词: Caenorhabditis elegans; RNA binding protein; RNA biosynthesis; SDS polyacrylamide gel electrophoresis; adenosinetriphosphatase; biochemistry; developmental genetics; enzyme activity; enzyme substrate; eukaryote; fluorescence microscopy; gel mobility shift assay; genetic regulation; genetic transcription; helicase; immunoprecipitation; messenger RNA; molecular cloning; protein protein interaction; protein structure function; thin layer chromatography

DESCRIPTION (provided by applicant): Organisms develop from reproductive cells in precisely organized patterns that are controlled by complex cellular mechanisms. Many of these mechanisms are conserved throughout higher eukaryotes. Reproductive cells are a type of stem cell that has the unique ability to divide to produce more stem cells or to divide and differentiate into a complete organism. The mechanisms regulating stem cell maintenance are not well understood, and they are important because stem cells have the potential to treat human diseases such as Parkinson's disease. This project involves a study of the regulation of stem cell proliferation in a model organism, the nematode C. elegans. These animals have a gonad containing stem cells, and the focus of this project is to determine the function of a specific protein controlling proliferation of these cells called RNA Helicase A (RHA-1). Helicases use energy from ATP hydrolysis to separate duplex nucleic acids into single strands, and RNA helicases are involved in various steps of mRNA metabolism, such as transcription and splicing. RNA helicase A is essential for proper development in mice, flies, and the model organism C. elegans. Experiments performed thus far demonstrate that RHA-1 is required for germline proliferation, but the mechanism by which RHA-1 controls proliferation is not known. A comparison of the phenotype of worms containing a large deletion in the rha-1 gene with the phenotype of other mutant worms with germline defects suggests that RHA-1 may act by controlling transcription or by participating in a signaling pathway in the germline. In this proposal, genetic and molecular biology experiments are presented to test whether RHA-1 has one of these functions or a different function. The biochemical activities of RHA-1 have not been determined, and these activities are presumably required for its role in germline proliferation. Thus, experiments are described to determine the biochemical activities of RHA-1 that are required for it to work properly. The regulation of cell division in the germline is also related to cancer treatment, since many processes controlling cell division are conserved, and these processes become disrupted in cancerous cells. The ultimate goal of this project is to determine how transcription and other cellular processes affect development in all eukaryotes, including humans.

描述（由申请人提供）：生物体由生殖细胞发育而成，生殖细胞具有精确的组织模式，由复杂的细胞机制控制。许多这些机制是保守的整个高等真核生物。生殖细胞是一种干细胞，它具有独特的分裂能力，可以产生更多的干细胞，或者分裂和分化成一个完整的生物体。调节干细胞维持的机制还不清楚，它们很重要，因为干细胞有治疗人类疾病如帕金森病的潜力。该项目涉及对模式生物--线虫C中干细胞增殖调节的研究。优雅的这些动物有一个含有干细胞的性腺，该项目的重点是确定控制这些细胞增殖的特定蛋白质的功能，称为RNA解旋酶A（RHA-1）。解旋酶使用来自ATP水解的能量将双链核酸分离成单链，并且RNA解旋酶参与mRNA代谢的各个步骤，例如转录和剪接。RNA解旋酶A是小鼠、苍蝇和模式生物C的正常发育所必需的。优美的迄今为止进行的实验表明，RHA-1是生殖系增殖所必需的，但RHA-1控制增殖的机制尚不清楚。在rha-1基因中含有大的缺失的蠕虫的表型与具有生殖系缺陷的其他突变体蠕虫的表型的比较表明，RHA-1可能通过控制转录或通过参与生殖系中的信号传导途径起作用。在这项提议中，提出了遗传和分子生物学实验来测试RHA-1是否具有这些功能之一或不同的功能。RHA-1的生物化学活性尚未确定，这些活性可能是其在生殖系增殖中的作用所必需的。因此，描述了确定RHA-1正常工作所需的生化活性的实验。生殖系中细胞分裂的调节也与癌症治疗有关，因为许多控制细胞分裂的过程是保守的，并且这些过程在癌细胞中被破坏。该项目的最终目标是确定转录和其他细胞过程如何影响包括人类在内的所有真核生物的发育。