E2F7 & E2F8 in the control of transcription and cellular proliferation

E2F7

• 批准号: 7749926
• 负责人: GUSTAVO Walter LEONE
• 金额: $ 31.13万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-10 至 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7749926
• 关键词: Ablation; Affinity Chromatography; Animals; Apoptosis; Binding; Biochemical; Biochemical Genetics; Biological; Cell Cycle; Cell Cycle Regulation; Cell Proliferation; Cells; Complex; Data; Defect; Development; Embryo; Family; Family member; Gene Expression; Gene Targeting; Generations; Genes; Genetic; Genetic Transcription; Homo; Individual; Laboratories; Macromolecular Complexes; Malignant Neoplasms; Methods; Multiprotein Complexes; Mus; Oncogenic; Pattern; Phenotype; Phosphotransferases; Play; Positioning Attribute; Protein Sequence Analysis; Proteins; Reagent; Recruitment Activity; Research; Role; Signal Pathway; Structure; System; Technology; Testing; Tissues; Transcription Repressor/Corepressor; Tumor Suppressor Proteins; Work; arm; base; combinatorial; in vivo; member; novel; overexpression; programs; promoter; protein complex; protein protein interaction; transcription factor

DESCRIPTION (provided by applicant): The E2F family of transcription factors is believed to play a critical role in the control of cellular proliferation. These factors are encoded by distinct genes and have both tumor suppressor and oncogenic functions (1-2). Our laboratory identified E2F7 and E2F8 as the final two members of this transcription factor family (5, 6). The preliminary data presented in this proposal highlight several unique features of these two E2Fs that place them in a subclass of their own. These salient features include their ability to form homodimers and heterodimers, to associate with a large cadre of transcriptional co-repressors, to silence gene expression, and to block cellular proliferation. While they lack a typical Rb-binding domain, E2F7 can specifically interact with Rb related proteins and can thus recruit E2F8 to Rb-containing complexes. As a result, the E2F7/8 arm of the E2F network remains under the control of the cycling dependent kinase (CDK) signaling pathway. The fact that E2F7 and E2F8 have an identical pattern of cell cycle dependent and tissue- specific expression, together with their ability to homo- and hetero-dimerize, raises the possibility that they may have both unique and shared functions in the animal. A multi-faceted effort in the laboratory has yielded key technical developments, including an affinity purification strategy to purify E2F7/8-associated proteins, promoter-array technologies to identify target genes, and gene targeting approaches to disrupt E2F7 and E2F8 in mice. These advances place our research group in a strong position to make significant advances towards a mechanistic understanding of how this important arm of the E2F family of factors controls the cell cycle and cell proliferation. The overarching hypothesis of this proposal is that E2F7 and E2F8 function as transcriptional repressors to negatively control cellular proliferation. Three specific aims utilizing biochemical, biophysical, global gene array, and genetic approaches will directly test this hypothesis: Specific Aim 1. To identify and characterize E2F7- and E2F8-associated macromolecular protein complexes. Specific Aim 2. To identify E2F7 and E2F8 transcriptional targets. Specific Aim 3. To determine the mechanism of E2F7 and E2F8 action in the control of transcription. This work will elucidate the individual and combinatorial contributions made by these two highly related family members towards the overall understanding of E2F transcriptional activity.

描述（由申请人提供）：E2F转录因子家族被认为在控制细胞增殖中起着至关重要的作用。这些因素由不同的基因编码，并具有肿瘤抑制和致癌功能（1-2）。我们的实验室将E2F7和E2F8确定为该转录因子家族的最后两个成员（5，6）。该提案中提供的初步数据突出了这两个E2F的几个独特功能，它们将它们置于自己的子类中。这些显着特征包括它们形成同二聚体和异二聚体的能力，与大量的转录共抑制剂联系在一起，使基因表达沉默并阻止细胞增殖。尽管它们缺乏典型的RB结合域，但E2F7可以特异性地与RB相关的蛋白质相互作用，因此可以募集E2F8到含RB的复合物。结果，E2F网络的E2F7/8 ARM保持在循环依赖激酶（CDK）信号通路的控制之下。 E2F7和E2F8具有相同的细胞循环依赖性和组织表达的模式，以及它们的同型和异性二聚体的能力，这增加了它们在动物中可能具有独特和共享功能的可能性。实验室中的多方面努力产生了关键的技术发展，包括纯化E2F7/8相关蛋白的亲和力纯化策略，启动子阵列技术以识别靶基因，以及破坏小鼠中E2F7和E2F8的基因靶向方法。这些进步使我们的研究小组处于强大的位置，以对E2F因子家族的这一重要部门如何控制细胞周期和细胞增殖，从而做出重大进展。该提案的总体假设是E2F7和E2F8充当转录阻遏物，可负控制细胞增殖。利用生化，生物物理，全球基因阵列和遗传方法的三个特定目的将直接检验以下假设：特定目的1。识别和表征E2F7和E2F8相关的大分子分子蛋白复合物。具体目标2。确定E2F7和E2F8转录靶标。具体目的3。确定转录控制中E2F7和E2F8作用的机制。这项工作将阐明这两个高度相关的家庭成员对E2F转录活动的整体理解所做的个体和组合贡献。