GENETIC STUDIES OF C-MYC GENE FUNTION IN THE CELL CYCLE

细胞周期中C-MYC基因功能的遗传学研究

• 批准号: 2181004
• 负责人: John M Sedivy
• 金额: $ 3.96万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-01-01 至 1995-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2181004
• 关键词: biological signal transduction; cell cycle; cell growth regulation; cyclins; gene expression; gene mutation; gene targeting; genetic markers; genetic promoter element; genetic recombination; genetic regulatory element; laboratory mouse; monoclonal antibody; neoplastic transformation; transcription factor; transposon /insertion element

DESCRIPTION (Adapted from Applicant's Abstract): An important part in unraveling the causes of cancer will be a detailed understanding of how mitogen-triggered signal transduction is integrated into the molecular mechanisms that govern the cell cycle. A large body of evidence indicates that the product of the c-myc gene is a transcription factor involved in these regulatory processes. Our knowledge of the role played by Myc is fragmentary in three important areas: 1) the mechanisms that regulate expression of the c- myc gene: 2) the downstream targets of Myc activity; and, 3) the mechanisms by which Myc regulates its targets. The goal of this proposal is a comprehensive investigation of signaling pathways both upstream and downstream of Myc. Emphasis is place on genetic experiments to understand how these regulatory processes operate in vivo. The first specific aim is to mutate promoter elements directly in a chromosomal c-myc gene. This will be accomplished using targeted homologous recombination, and will allow the in vivo study of specific regulatory circuits in the regulation of the c-myc gene. The second specific aim is to identify downstream Myc target genes, and to determine how Myc regulates their expression. This will be accomplished by testing the expression of specific putative targets in cells devoid of Myc activity (Myc- cells). A second approach will be to use a general screening method, differential display, to search for novel targets whose expression is altered in Myc- cells. Follow up experiments will identify the cis-acting elements in the targets genes that mediate the action of Myc. These experiments will include an investigating of how Myc regulates the expression of cyclins E and A, which have been implicated as downstream targets. The third specific aim is to disrupt the cyclin E gene, which will be accomplished by gene targeting. The comparison of Myc- and cyclin E- phenotypes will reveal which aspects of the growth regulatory activity of Myc are mediated by cyclin E. Experiments to further improve and generalize gene targeting methodologies are also a part of this specific aim. All the proposed experiments are founded on previous accomplishments of this laboratory: 1) the development of a new and efficient gene targeting strategy; 2) the isolation of Myc- cell lines; and, 3) the identification of cyclins E and A as Myc targets. Taken together, the experiments outlined in this proposal are aimed at providing a better understanding of the function of Myc in normal cellular physiology, and ultimately, its oncogenic action.

描述（改编自申请人的摘要）： 阐明癌症的原因将是对如何的详细理解 有丝分裂原触发的信号转导整合到分子中 控制细胞周期的机制。大量证据 表示C-MYC基因的乘积是转录因子 参与这些监管过程。我们对角色的了解 MYC在三个重要领域中扮演的是零碎的：1）机制 调节C- MYC基因的表达：2）下游目标 MYC活动； 3）MYC调节其机制 目标。该提案的目的是对 MYC上游和下游的信号通路。重点是 放在遗传实验上，以了解这些调节 过程在体内运行。 第一个具体目的是直接在A中突变启动子元素 染色体C-MYC基因。这将使用针对性 同源重组，并将允许对特定的体内研究 C-MYC基因调节中的调节回路。 第二个具体目的是识别下游MYC目标基因，并 确定MYC如何调节其表达。这将是 通过测试特定推定目标的表达来完成 缺乏MYC活性（MYC细胞）的细胞。第二种方法将是 要使用一般筛选方法，差分显示，以搜索 在Myc细胞中表达改变的新靶标。跟进 实验将确定目标基因中的顺式作用元素 调解了MYC的动作。这些实验将包括 调查MYC如何调节细胞周期蛋白E和A的表达 这与下游目标有关。 第三个具体目的是破坏细胞周期蛋白基因，这将是 通过基因靶向完成。 Myc-和Cyclin E-的比较 表型将揭示生长调节活动的哪些方面 MYC的介导了Cyclin E.实验以进一步改进和 概括基因靶向方法也是该特定的一部分 目的。 所有提出的实验都是基于以前的成就 该实验室：1）开发新的高效基因 目标策略； 2）分离Myc-细胞系； 3） 鉴定细胞周期蛋白E和A作为MYC靶标。总的来说， 该提案中概述的实验旨在提供更好的 了解MYC在正常细胞生理学中的功能，以及 最终，其致癌作用。