Functional Studies of the CDK-module of the Human Mediator Complex

人类介导复合体 CDK 模块的功能研究

• 批准号: 0842974
• 负责人: Joaquin Espinosa
• 金额: $ 45.4万
• 依托单位: University of Colorado at Boulder
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-01 至 2013-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0842974&HistoricalAwards=false
• 关键词: Functional; Studies; CDK; module; Human

Intellectual merit. A major challenge in modern biology is to elucidate the molecular mechanisms employed by cells to regulate expression of thousands of genes in a tissue-, signaling-, and development-specific manner. Proper regulation of gene activity is a key element of every major biological process. Specialized protein machineries can turn genes ON and OFF in a tightly regulated manner. One such protein complex, known as Mediator, is recognized as a critical regulator of gene activity from yeast to humans, yet little is known about its mechanism of action. This research project investigates a four-protein sub-complex of Mediator known as the CDK-module, which can function to either activate or repress genes. The goal of this project is to elucidate the molecular mechanism by which the CDK module activates genes including the regulatory steps at which it acts, defining the requirement for different domains of the catalytically active subunit, and delineating the roles of the other subunits. Building on the identification of specific genes in mammalian cells that are clearly regulated by the CDK-module, sophisticated biochemical and molecular biology techniques will be employed to manipulate the activity of this complex inside cells and thus obtain much needed mechanistic information. This research project will greatly improve our understanding of Mediator in particular and the evolution of gene expression control mechanisms in general. Given the quasi-universal role of Mediator in the control of gene expression, the discoveries arising from this project will have far reaching implications in the fields of cell and molecular biology. Moreover, the innovative experimental approaches not previously utilized to study Mediator in vertebrate cells will almost certainly provide novel insights. Broader impacts. The activities in this project will be carried out by a teaching-research team that strongly promotes science education and diversity in science. Preliminary results have been obtained by partnerships of undergraduate and graduate students working in a laboratory with an overall 1:1 female to male ratio and 1/3 of the lab members representing ethnic minorities. This grant will fund one such undergraduate-graduate student collaboration. Both the host institution and the Principal Investigator actively participate in programs funded by NSF and other agencies that aim to: a) increase the number of minority students earning PhDs, b) provide hands-on research experience to undergrads, c) prepare undergraduate students to pursue PhDs in the biosciences, and d) increase the representation of women in leadership positions in science and engineering. The success of these endeavors is demonstrated by the fact that every scientific publication from this research group has one or more undergraduate authors, and that several of the past undergraduate trainees are now enrolled in PhD programs. The Principal Investigator, Dr. Espinosa, is an active member of the local Hispanic community and a recognized science educator at CU-Boulder, where he is the instructor of a ~150-student undergraduate course and co-instructor in the graduate program of his home Department. He teaches topics related to his research at both venues. This grant will help to cement a culture of integrated research and education within this burgeoning group, as well as to promote diversity at the host institution.

智力上的优点。现代生物学的一个主要挑战是阐明细胞以组织特异性、信号特异性和发育特异性方式调节数千个基因表达的分子机制。基因活性的适当调节是每个主要生物过程的关键因素。专门的蛋白质机器可以以严格调控的方式打开和关闭基因。一种这样的蛋白质复合物，称为介体，被认为是从酵母到人类的基因活性的关键调节因子，但对其作用机制知之甚少。该研究项目研究了一种称为CDK模块的四蛋白亚复合物，它可以激活或抑制基因。 该项目的目标是阐明CDK模块激活基因的分子机制，包括其作用的调节步骤，定义催化活性亚基的不同结构域的要求，并描绘其他亚基的作用。 在哺乳动物细胞中明确受CDK模块调控的特定基因的鉴定的基础上，将采用复杂的生物化学和分子生物学技术来操纵细胞内这种复合物的活性，从而获得急需的机制信息。这个研究项目将极大地提高我们对Mediator的理解，特别是对基因表达控制机制的进化的理解。鉴于Mediator在控制基因表达中的准普遍作用，该项目的发现将在细胞和分子生物学领域产生深远的影响。此外，以前没有用于研究脊椎动物细胞中的介体的创新实验方法几乎肯定会提供新的见解。更广泛的影响。该项目的活动将由一个大力促进科学教育和科学多样性的教学研究小组开展。通过在一个男女比例为1：1的实验室工作的本科生和研究生的伙伴关系取得了初步结果，实验室成员中有1/3是少数民族。这笔赠款将资助一个这样的本科生-研究生合作。主办机构和主要研究者都积极参与由NSF和其他机构资助的项目，这些项目旨在：a）增加获得博士学位的少数民族学生的数量，B）为本科生提供实践研究经验，c）为本科生攻读生物科学博士学位做好准备，d）增加女性在科学和工程领导职位中的代表性。这些努力的成功是由这个研究小组的每一个科学出版物都有一个或多个本科作者的事实证明的，而且过去的几个本科学员现在都在攻读博士学位。 主要研究者埃斯皮诺萨博士是当地西班牙裔社区的活跃成员，也是CU-博尔德公认的科学教育家，他是一门约150名学生的本科课程的讲师，也是他所在系研究生课程的共同讲师。他在两个地点教授与他的研究有关的主题。这笔赠款将有助于巩固这一新兴群体中的综合研究和教育文化，并促进东道机构的多样性。