MOLECULAR GENETICS OF CHROMATID SEPARATION--ROLE OF BRRN

描述 （摘自申请人摘要）本提案旨在适应 Olga A博士的培训和职业发展目标。卡贝罗 的 候选人接受过心血管生理学和分子生物学的研究生培训 生物物理学和细胞生物学和分子生物学博士后培训 遗传学 职业发展计划（CDP）将为她提供 在蛋白质生物化学，细胞生物学技术和 转基因动物的产生。 Cabello博士最近被任命为 她的长期职业目标是建立一个 学术研究计划侧重于哺乳动物有丝分裂的分子分析 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 研究活动最初将在 H博士的实验室Youssoufian，分子和人类系 遗传学 随后，候选人将占据自己的研究空间， 细胞生物学系，在B博士的密切监督下。R. 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：新复制的姐妹染色单体的邻接机制 臂，并允许他们在后期精确分离是不完全的 可以理解，但可能涉及多个过程，包括降解 粘附蛋白和拓扑异构酶II（topo II）的作用。 她 建议分析最近描述的人类和小鼠同源物， 果蝇蛋白不育。 染色单体臂分离需要贫瘠 可能是因为其对Topo II活性的调节作用。 他们有 部分表征了编码人类不育基因（BRRN-1）， 确定了这个假定基因家族的其他成员。 他们打算测试 假设贫瘠的功能是保守的， 哺乳动物细胞中观察到的类似于果蝇。 他们会采用 研究BRRN-1的功能及其与 在人类细胞中的作用机制a）它们将确定贫瘠-1 表达和细胞周期依赖性亚细胞定位，B）它们将 然后评估潜在的结构和功能相互作用， barn-1与topo II和SMC的哺乳动物同源物（结构 维持染色体）蛋白质。 他们将分析 小鼠胚胎干细胞中mbrrn 1基因的简单和条件性靶向失活 细胞 在成功生产这些细胞系后，他们将尝试 杂合突变小鼠的生产。 拟议的研究应 产生关于拓扑异构酶II的调节及其 参与哺乳动物细胞的后期过程。 此外，他们可能 为人类基因组稳定性疾病提供了新的见解。 (End的 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)