The E2F1 Post-Translational Modification Code

E2F1 翻译后修饰代码

• 批准号: 10440288
• 负责人: David G. Johnson
• 金额: $ 43.55万
• 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 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10440288
• 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; 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.

项目概要 在大多数癌症中，E2F1 转录因子通过 p16INK4A-细胞周期蛋白 D-RB 的破坏而失调 肿瘤抑制途径，但 E2F1 在癌症中发挥的作用很复杂，因为它可以促进或抑制 肿瘤的发展取决于具体情况。 E2F1 还可以通过以下方式在分子水平上具有相反的活性： 通过促进增殖或凋亡在细胞水平上激活或抑制转录。而且， E2F1 可以直接刺激 DNA 修复，与其在转录中的作用无关。这些的长期目标 研究的目的是了解E2F1的各种功能在细胞中是如何调节的，以及每种功能是如何调节的 活性有助于癌症的发展或抑制。 E2F1 被磷酸化、乙酰化、 并因 DNA 损伤而甲基化，但这些修饰如何改变 E2F1 的活性仍然未知 不太了解。该提议基于以下假设：翻译后修饰 E2F1 由相互作用的蛋白质读取，以调节 E2F1 定位和 DNA 修复功能， 转录和肿瘤发生。 TopBP1 蛋白在磷酸化后与 E2F1 特异性结合 通过 ATM/ATR 激酶，这种相互作用导致 E2F1 募集到 DNA 损伤位点。目标1 研究将使用一种新型敲入小鼠模型（S29A）来阻断 E2F1 磷酸化来确定 E2F1及其相关蛋白在动态修饰DNA位点染色质结构中的功能 损坏以利维修。初步数据表明 E2F1 乙酰化为 CBP 和 p300 乙酰转移酶的溴结构域。一种阻断 E2F1 的新型敲入小鼠模型 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 修改 DNA 损伤位点的染色质结构以促进修复。这些敲入小鼠也将被使用 检查 E2F1 乙酰化在调节转录和抑制肿瘤发展中的作用（目标 2）。 TDRD3 蛋白被鉴定为 E2F1 精氨酸甲基化标记 (R109me) 的潜在读取器。这 将研究 TDRD3 在调节 DNA 修复和转录中 E2F1 依赖性功能中的作用，如果 如有必要，将开发敲入小鼠模型来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 翻译后修饰代码是如何被读取的 调节 DNA 修复和转录的各种染色质修饰活性。确定如何这些 E2F1翻译后修饰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治疗效果。

项目成果

The E2F1 transcription factor and RB tumor suppressor moonlight as DNA repair factors.

• DOI: 10.1080/15384101.2020.1801190
• 发表时间: 2020-09
• 期刊: Cell cycle (Georgetown, Tex.)
• 作者: Manickavinayaham S;Velez-Cruz R;Biswas AK;Chen J;Guo R;Johnson DG
• 通讯作者: Johnson DG

Direct Regulation of DNA Repair by E2F and RB in Mammals and Plants: Core Function or Convergent Evolution?

• DOI: 10.3390/cancers13050934
• 发表时间: 2021-02-24
• 期刊: Cancers
• 影响因子: 5.2
• 作者: Manickavinayaham S;Dennehey BK;Johnson DG
• 通讯作者: Johnson DG