CAREER: Nuclear RNA Processing, Coilin and Cajal Bodies

职业：核 RNA 处理、Coilin 和 Cajal 体

• 批准号: 0344066
• 负责人: Michel Bellini
• 金额: $ 72万
• 依托单位: University of Illinois at Urbana-Champaign
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-15 至 2010-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0344066&HistoricalAwards=false
• 关键词: CAREER; Nuclear; RNA; Processing; Coilin

How molecular events, such as RNA transcription and maturation, influence nuclear architecture is an important physiological aspect of the eukaryotic cell nucleus. In recent years this central question promoted renewed interest in the organization of the nucleus and rediscovery of several discrete nuclear organelles. The overall objective of this research project is to evaluate the role of one of these nuclear organelles, the Cajal body (CB), in the regulation of nuclear RNA processing. The main experimental system used is the nucleus of Xenopus laevis oocytes, because it provides the unique opportunity to approach this question with unequaled spatial resolution. Two essential questions will be addressed: What is the functional relevance of small nuclear ribonucleoprotein particles (snRNPs) trafficking through CBs, and how does coilin, the signature protein of CBs, recruit snRNPs to CBs? Coilin will be used as a molecular tool to prevent the entry of snRNPs into CBs, without affecting the apparent physiology of the latter. It will allow the investigation in vivo of whether the flow of snRNPs through CBs is required for some aspects of their maturation and/or the processing of pre-messenger RNAs. Biochemical fractionations will also be performed on oocyte extracts to identify the putative nucleoplasmic proteins and RNAs that interact with coilin. This information will then be used in various cell systems to define the molecular mechanisms by which coilin, and the newly identified factors, recruit snRNPs to CBs. Investigation of coilin's biochemistry and cellular dynamics in relation with snRNPs will provide new insights into the nuclear roles of CBs. Because of its visual appeal and accessibility, this research is highly amenable to an active outreach program. It will be applied at several educational levels by mentoring a high school biology teacher and class, and introducing many undergraduate and community college students, including women and minorities, to the wonders of biology and new imaging technologies.

分子事件，如RNA转录和成熟，如何影响核结构是真核细胞核的一个重要生理方面。 近年来，这个中心问题促进了对细胞核组织的重新兴趣，并重新发现了几个离散的核细胞器。 本研究项目的总体目标是评估这些核细胞器之一，卡哈尔体（CB），在调节核RNA加工的作用。 使用的主要实验系统是非洲爪蟾卵母细胞的细胞核，因为它提供了独特的机会，以解决这个问题与无与伦比的空间分辨率。 两个基本问题将得到解决：什么是功能相关的小核核糖核蛋白颗粒（snRNP）贩运通过CB，以及如何coilin，CB的签名蛋白，招聘snRNP CB？ 卷曲蛋白将被用作防止snRNP进入CB的分子工具，而不影响后者的表观生理学。 它将允许在体内调查snRNP通过CB的流动是否是其成熟和/或前信使RNA加工的某些方面所需的。 还将对卵母细胞提取物进行生化分级，以鉴定与卷曲蛋白相互作用的推定核质蛋白和RNA。 然后，这些信息将用于各种细胞系统，以确定卷曲蛋白和新鉴定的因子将snRNP招募到CB的分子机制。 卷曲蛋白的生物化学和细胞动力学与snRNP的调查将提供新的见解CBs的核作用。 由于其视觉吸引力和可访问性，这项研究是非常适合一个积极的推广计划。 它将通过指导高中生物教师和班级，并向包括妇女和少数民族在内的许多本科生和社区学院学生介绍生物学和新成像技术的奇迹，在几个教育层次上应用。