The E2F1 Post-Translational Modification Code

E2F1 翻译后修饰代码

• 批准号: 10218068
• 负责人: David G. Johnson
• 金额: $ 44.44万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10218068
• 关键词: Acetylation; Active Sites; Apoptosis; Arginine; Binding; Bromodomain; CDKN2A gene; Cell Fate Control; Cell Proliferation; Cell physiology; Cells; ChIP-seq; Chromatin; Chromatin Structure; Code; Complex; Cyclin D1; DNA; DNA Binding Domain; DNA Damage; DNA Repair; Data; Defect; Development; Double Strand Break Repair; E2F transcription factors; EP300 gene; Effectiveness; Environment; Enzymes; Gene Expression; Genes; Genetic Transcription; Genome Stability; Goals; Growth; Histone Acetylation; Histone H3; Human; Hypersensitivity; Impairment; Ionizing radiation; Kinetics; Knock-in; Knock-in Mouse; Laboratories; Lysine; Malignant Neoplasms; Methylation; Modeling; Modification; Molecular; Monitor; Mus; Mutation; Nucleosomes; Pathway interactions; Phosphorylation; Phosphotransferases; Play; Post-Translational Protein Processing; Predisposition; Proteins; Publishing; Reader; Role; SMARCA4 gene; Signal Transduction; Site; Skin Carcinogenesis; System; Testing; Therapeutic Intervention; Topoisomerase; Tumor Suppressor Proteins; UV induced; UV induced DNA damage; Ultraviolet Rays; base; cancer cell; chromatin remodeling; endonuclease; experimental study; histone acetyltransferase; in vivo; inhibitor/antagonist; mouse model; novel; novel therapeutics; p300/CBP-Associated Factor; prevent; promoter; recruit; repaired; response; tool development; tumor; tumor growth; tumorigenesis

PROJECT SUMMARY The E2F1 transcription factor is deregulated in most cancers through disruptions in the p16INK4A-cyclin D-RB tumor suppressor pathway but the role E2F1 plays in cancer is complex as it can either promote or inhibit tumor development depending on the context. E2F1 can also have opposing activities at the molecular level by activating or repressing transcription and at the cellular level by promoting proliferation or apoptosis. Moreover, E2F1 can directly stimulate DNA repair independent of its role in transcription. A long-term goal of these studies is to understand how the various functions of E2F1 are regulated in the cell and how each activity contributes to the development or suppression of cancer. E2F1 is phosphorylated, acetylated, and methylated in response to DNA damage but how these modifications alter the activities of E2F1 remains poorly understood. This proposal is based on the hypothesis that post-translational modifications on E2F1 are read by interacting proteins to regulate E2F1 localization and functions in DNA repair, transcription and tumorigenesis. The TopBP1 protein specifically binds to E2F1 when it is phosphorylated by the ATM/ATR kinases and this interaction results in the recruitment of E2F1 to sites of DNA damage. Aim 1 studies will use a novel knock-in mouse model (S29A) that blocks E2F1 phosphorylation to determine the function of E2F1 and its associated proteins in dynamically modifying chromatin structure at sites of DNA damage to facilitate repair. Preliminary data demonstrates that E2F1 acetylation creates a binding motif for the bromodomains of the CBP and p300 acetyltransferases. A novel knock-in mouse model that blocks E2F1 acetylation will be used to study how the E2F1-CBP/p300 interaction regulates histone acetylation and modifies chromatin structure at sites of DNA damage to facilitate repair. These knock-in mice will also be used to examine the role of E2F1 acetylation in regulating transcription and suppressing tumor development (Aim 2). The TDRD3 protein was identified as a potential reader of an E2F1 arginine methylation mark (R109me). The role of TDRD3 in regulating E2F1-dependent functions in DNA repair and transcription will be studied and, if warranted, a knock-in mouse model will be developed to establish the role of this modification in modulating tumorigenesis. Together, these studies will reveal how the E2F1 post-translational modification code is read by various chromatin-modifying activities to regulate both DNA repair and transcription. Determining how these E2F1 post-translational modifications impact the development of cancer will reveal new opportunities to modulate the activities of E2F1 to inhibit tumor growth and enhance the effectiveness of therapies.

项目摘要 通过p16ink4a-cyclin d-rb，大多数癌症中E2F1转录因子在大多数癌症中受到了管制。 肿瘤抑制途径，但E2F1在癌症中的作用很复杂，因为它可以促进或抑制 肿瘤发展取决于上下文。 E2F1也可以通过分子水平进行相反的活动 通过促进增殖或凋亡来激活或抑制转录水平。而且， E2F1可以直接刺激DNA修复，而与其在转录中的作用无关。这些的长期目标 研究是为了了解如何在细胞中调节E2F1的各种功能以及如何调节每个功能 活动有助于癌症的发展或抑制。 E2F1是磷酸化的，乙酰化的， 并响应DNA损伤而甲基化，但是这些修饰如何改变E2F1的活性。 理解不佳。该提议基于以下假设。 通过相互作用蛋白来调节E2F1定位和DNA修复中的功能，读取E2F1， 转录和肿瘤发生。 TOPBP1蛋白在磷酸化时特异性与E2F1结合 通过ATM/ATR激酶和这种相互作用导致E2F1募集到DNA损伤部位。目标1 研究将使用一种新型的敲入小鼠模型（S29A），该模型阻止E2F1磷酸化来确定 E2F1及其相关蛋白在动态修饰DNA位点的染色质结构中的功能 损坏促进维修。初步数据表明，e2f1乙酰化为 CBP和p300乙酰转移酶的溴结构域。阻止E2F1的新型敲入鼠标模型 乙酰化将用于研究E2F1-CBP/P300相互作用如何调节组蛋白乙酰化和 修饰DNA损伤部位的染色质结构，以促进修复。这些敲门鼠也将使用 检查E2F1乙酰化在调节转录和抑制肿瘤发育中的作用（AIM 2）。 TDRD3蛋白被确定为E2F1精氨酸甲基化标记（R109ME）的潜在读取器。这 将研究TDRD3在调节E2F1依赖性功能中的作用在DNA修复和转录中的作用，如果 有保证，将开发一个敲入鼠标模型，以确定这种修饰在调节中的作用 肿瘤发生。这些研究将共同​​揭示E2F1翻译后修改代码如何通过 各种染色质修改活性，以调节DNA修复和转录。确定这些方式 E2F1翻译后修饰会影响癌症的发展，将揭示新的机会 调节E2F1的活性以抑制肿瘤生长并增强疗法的有效性。