XRCC4 AND V<D>J RECOMBINATION AND DNA REPAIR

XRCC4 和 V<D>J 重组和 DNA 修复

• 批准号: 2882102
• 负责人: KAREN M FRANK
• 金额: $ 7.76万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-03-01 至 2002-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2882102
• 关键词: DNA binding protein; DNA repair; animal genetic material tag; biochemical evolution; cell differentiation; gene complementation; gene mutation; genetic recombination; laboratory mouse; lymphocyte; phosphorylation; protein structure function; site directed mutagenesis; tissue /cell culture; yeast two hybrid system

DESCRIPTION: a. Candidate: Dr. Karen M. Frank received her M.D. and Ph.D. degrees from the University of Pennsylvania in 1994. She has been a clinical pathology resident and research fellow at Brigham and Women's Hospital and Children's Hospital since 1994. She served as Chief Resident in clinical pathology at Brigham and Women's Hospital from January 1996 to June 1996. Dr. Frank listed six published papers, in which she was first author on four. The candidate's goal is to perform immunology research at a major medical center with an academic faculty appointment, and with a joint appointment in a tertiary care hospital. b. Mentor: Dr. Frederick Alt is the Charles Janeway Professor of Pediatrics and Professor of Genetics and Pediatrics at the Harvard Medical School, and an Investigator of the Howard Hughes Medical Institute at the Children's Hospital. c. Career Development Plan: The candidate will devote 85-90 percent time to research. She will learn new techniques in molecular biology, increase her knowledge of immunology, and develop skills in experimental design and presentation of research results. A modest amount of time will be devoted to teaching and to learning to manage a laboratory. d. Research Plan: Dr. Frank proposes to investigate the function of the newly cloned gene XRCC4 in the V(D)J recombination process. XRCC4 was isolated in the mentor's laboratory on the basis of its potential involvement in V(D)J recombination and double stranded (DS) DNA break repair. The hypothesis of the project is that the XRCC4 protein plays a direct role in the V(D)J recombination process. Three approaches will be used to address the hypothesis. In Specific Aim 1, Dr. Frank will attempt to determine functional domains of XRCC4 by mutational analysis and by examining the evolutionary conservation of amino acid sequence. Dr. Frank will also try to determine if XRCC4 is phosphorylated and whether phosphorylation has any role in its function. In Specific Aim 2, Dr. Frank proposes to investigate whether XRCC4 associates with known V(D)J recombination components such as RAG-1, RAG-2, Ku 80, Ku 70 and DNA-PKCS and other newly discovered proteins. In Specific Aim 3, Dr. Frank will generate XRCC4 knockout (KO) mice and study the role of XRCC4 in lymphocyte development and in DNA repair. e. Environment and Institutional Support: The training will take place in Dr. Alt's laboratory in the Enders Research Building of the Children's Hospital.

描述： 一个。候选人：Karen M. Frank 博士获得医学博士学位和博士学位。度数从 1994年在宾夕法尼亚大学获得临床病理学博士学位 布莱根妇女医院和儿童医院住院医师和研究员 自 1994 年起入院。她曾担任以下医院临床病理科的首席住院医师： 1996 年 1 月至 1996 年 6 月在布莱根妇女医院工作。 Frank 博士 列出了六篇已发表的论文，其中四篇是她的第一作者。 这 候选人的目标是在大型医疗中心进行免疫学研究 具有学术教员任命，并在一个联合任命 三级甲等医院。 b.导师：Frederick Alt 博士是 Charles Janeway 儿科教授 哈佛医学院遗传学和儿科教授，以及 儿童医院霍华德休斯医学研究所的研究员 医院。 c.职业发展计划：候选人将投入 85-90% 的时间 研究。 她将学习分子生物学的新技术，提高她的能力 免疫学知识，并培养实验设计和 研究成果的介绍。 将投入适量的时间 教学和学习管理实验室。 d.研究计划：弗兰克博士提议研究 V(D)J重组过程中新克隆的基因XRCC4。 XRCC4 是 根据其潜力在导师的实验室中隔离 参与 V(D)J 重组和双链 (DS) DNA 断裂 维修。 该项目的假设是XRCC4蛋白发挥直接作用 在V(D)J重组过程中。 将使用三种方法 解决假设。 在具体目标 1 中，弗兰克博士将尝试 通过突变分析确定 XRCC4 的功能域 检查氨基酸序列的进化保守性。 弗兰克博士 还将尝试确定 XRCC4 是否被磷酸化以及是否 磷酸化对其功能有任何作用。 在具体目标 2 中，弗兰克博士 提议调查 XRCC4 是否与已知的 V(D)J 相关 重组成分，例如 RAG-1、RAG-2、Ku 80、Ku 70 和 DNA-PKCS 以及 其他新发现的蛋白质。 在特定目标 3 中，弗兰克博士将生成 XRCC4敲除（KO）小鼠并研究XRCC4在淋巴细胞中的作用 发育和 DNA 修复。 e.环境和机构支持：培训将在 阿尔特博士位于儿童医院恩德斯研究大楼的实验室 医院。