MOLECULAR GENETICS OF CHROMATID SEPARATION--ROLE OF BRRN

描述 (改编自申请人的摘要)这项建议是为适应 Olga A.Cabello博士的培训和职业发展目标。这个 有心血管生理学和分子专业研究生学历 生物物理学，细胞生物学和分子博士后培训 遗传学。拟议的职业发展计划(CDP)将为她提供 蛋白质生物化学、细胞生物学技术和 转基因动物的产生。卡贝罗博士最近被任命为一家 初级教员职位，她的长期职业目标是建立一个 专注于哺乳动物有丝分裂的分子分析的学术研究计划 事件及其在基因组稳定性中的作用。环境：CDP是 分为两个阶段，围绕研究的发展而设计 计划一下。初步的研究活动将在 优素菲安博士实验室，分子与人类系 遗传学。随后，候选人将占据她自己的研究空间 在B.R.博士的密切监督下，细胞生物系。 布林克利，确保成功地逐步过渡到学术独立。 五年期CDP的所有方面都将由一个咨询委员会指导。 研究计划：与新复制的姐妹染色单体相邻的机制 手臂，并允许它们在后期精确分离是不完全的 了解，但可能涉及多个过程，包括降级 黏附蛋白与拓扑毒物异构酶II的作用。她 建议分析最近描述的人和老鼠的同源基因 果蝇蛋白质贫乏。染色单体臂分离需要不育系 可能是由于其对Topo II活性的调节作用。他们有 部分鉴定了编码人类不育(BRRN-1)的基因，并具有 确定了这个假定的基因家族的其他成员。他们打算测试 不育症的功能是保守的，其活动在 哺乳动物细胞与在果蝇中观察到的细胞相似。他们将雇用 了解BRRN-1及其功能的几种策略 在人类细胞中的作用机制a)它们将决定Branen-1 表达和依赖于细胞周期的亚细胞定位，b)它们将 然后评估潜在的结构和功能相互作用 含有Topo II和SMC的哺乳动物同源物的Branren-1(结构 维持染色体)蛋白质。他们将分析 小鼠mbrrn1在培养ES中的简单和有条件的靶向失活 细胞。在成功生产这些细胞系后，他们将尝试 杂合子突变小鼠的产生。建议的研究应 获得关于拓扑异构酶II及其调控的有价值的信息 参与哺乳动物细胞的后期过程。此外，他们还可以 为人类基因组稳定性紊乱提供了新的见解。(末尾 摘要)

DESCRIPTION (Adapted from applicant's abstract) This proposal has been designed to suit the training and career development objectives of Dr. Olga A. Cabello. The candidate has graduate training in Cardiovascular Physiology and Molecular Biophysics, and postdoctoral training in Cell Biology and Molecular Genetics. The proposed Career Development Plan (CDP) will provide her with additional skills in protein biochemistry, cell biological techniques and generation of transgenic animals. Dr. Cabello was recently appointed to a junior faculty position and her long term career goal is to establish an academic research program focused on molecular analysis of mammalian mitotic events and their role in genomic stability. Environment: The CDP is divided in two phases and is designed around the development of the Research Plan. Initially, the research activities will be conducted in the laboratory of Dr. H. Youssoufian, Department of Molecular and Human Genetics. Subsequently, the candidate will occupy her own research space in the Department of Cell Biology, under close supervision of Dr. B. R. Brinkley, ensuring a successful gradual transition to academic independence. All aspects of the five year CDP will be guided by an Advisory Committee. Research Plan: the mechanisms that adjoin newly replicated sister chromatid arms, and permit their precise separation at anaphase are incompletely understood, but may involve multiple processes including the degradation of adhesion proteins and the action of toppoisomerase II (topo II). She propose to analyze the human and mouse homologues of the recently describe Drosophila protein barren. Barren is required for chromatid arm separation possibly because of its modulatory action on topo II activity. They have partially characterized the gene encoding human barren (BRRN-1) and have identified other members of this putative gene family. They intend to test the hypothesis that barren function is conserved and that its activity in mammalian cells is similar to that observed in Drosophila. They will employ several strategies to gain insight into the function of BRRN-1 and its mechanism of action in human cells a) they will determine barren-1 expression and cell cycle-dependent subcellular localization, b) they will then evaluate the potential structural and functional interaction of barren-1 with topo II and mammalian homologues of SMC (structural maintenance of chromosomes) proteins. They will analyze the effect of simple and conditional targeted inactivation of mouse mbrrn1 in cultured ES cells. After successful production of these cell lines they will attempt the production of heterozygous mutant mice. The proposed studies should yield valuable information on the regulation of topoisomerase II and its participation in the anaphase process in mammalian cells. Also, they may provide new insights into human disorders of genomic stability. (End of Abstract)