ROLE OF E2F IN REGULATION OF CELL CYCLE AND TUMOR DEVELOPMENT

E2F 在调节细胞周期和肿瘤发展中的作用

• 批准号: 6300269
• 负责人: Jacqueline A. Lees
• 金额: $ 13.53万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-01 至 2001-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6300269
• 关键词: cell cycle; gene mutation; gene targeting; genetic transcription; laboratory mouse; neoplasm /cancer genetics; retinoblastoma protein; tissue /cell culture; transcription factor; tumor suppressor genes

The developmental and tumor-suppressive properties of the retinoblastoma protein (pRB) both appear to be at least partially dependent upon its ability to regulate the transcriptional activity of a discrete subset of the E2F transcription factors. Animal models will be used to study the developmental role of the pRB-specific E2Fs and determine how their regulation contributes to the properties of the retinoblastoma protein. Novel mouse strains with germline mutations in the E2F-2 and E2F-3 genes will be created using standard gene targeting technology. Analysis of these mice will establish how loss of either single or multiple pRB- specific E2Fs affects the development of embryonic and adult mice. Crossbreeding will then be used to determine whether mutation of the these genes is sufficient to block either the tumor formation that occurs in the Rb+/- mice or the widespread apoptosis that occurs in the Rb-/- embryos. This will help to establish how E2F-regulation contributes to the developmental and tumor-suppressive properties of pRB. The effect of these mutations will also be examined at the molecular level using the single and compound mutant mouse strains to generate primary cells lines. In each case, the spectrum of E2F complexes, the degree and timing of activation of E2F responsive genes and growth properties of these mutant cells will be analyzed. These studies will help to link the pRB-specific E2Fs to the activation of specific target genes and determine how these individual events contribute to growth stimulatory properties of the endogenous E2F. Finally, we will continue to develop in vitro transcription assays that will allow us to study the direct consequences of the interaction between pRB and its target E2Fs. In particular, this system will be optimized to study the mechanism by which pRB abolishes the basal transcription of promoters to which it is bound. The goal of these in vitro studies is to separate the E2F-specific and global repression properties of pRB, to establish the molecular mechanisms by which these effects are mediated, and to determine how each one contributes to the growth suppressive properties of this protein.

视网膜母细胞瘤的发育和肿瘤抑制特性 蛋白质（pRB）似乎都至少部分依赖于其 调节离散子集转录活性的能力 E2F转录因子。 动物模型将用于研究 pRB 特异性 E2F 的发育作用并确定它们如何 调节有助于视网膜母细胞瘤蛋白的特性。 E2F-2 和 E2F-3 基因发生种系突变的新型小鼠品系 将使用标准基因靶向技术创建。 分析 这些小鼠将确定单个或多个 pRB- 的丢失如何 特定的 E2F 影响胚胎和成年小鼠的发育。 然后将使用杂交来确定这些物种是否发生突变 基因足以阻止肿瘤的形成 Rb+/- 小鼠或 Rb-/- 胚胎中发生的广泛细胞凋亡。 这将有助于确定 E2F 监管如何促进 pRB 的发育和肿瘤抑制特性。 的效果 这些突变也将在分子水平上进行检查 单一和复合突变小鼠品系以产生原代细胞系。 在每种情况下，E2F 复合体的频谱、程度和时间 E2F 响应基因的激活和这些突变体的生长特性 将分析细胞。 这些研究将有助于将 pRB 特异性联系起来 E2Fs 来激活特定的靶基因并确定这些基因如何被激活 个别事件有助于促进生长的特性 内源性E2F。 最后我们会继续进行体外开发 转录测定将使我们能够研究直接后果 pRB 与其目标 E2F 之间的相互作用。 特别是，这 优化体系，研究PRB废止机制 它所结合的启动子的基础转录。 这些的目标 体外研究的目的是分离 E2F 特异性抑制和全局抑制 pRB 的特性，以建立这些分子机制 影响是中介的，并确定每一种影响如何贡献 该蛋白质的生长抑制特性。