BET regulation of mitosis and pluripotency establishment

有丝分裂和多能性建立的 BET 调节

• 批准号: 10455668
• 负责人: Kejin Hu
• 金额: $ 12.66万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 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 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10455668
• 关键词: Acetylation; Affinity; BRD2 gene; Binding; Biochemical Process; Bioinformatics; Biological; Biological Assay; Biological Process; Bromodomain; C-terminal; Cell Cycle; Cell Proliferation; Cell physiology; Cells; ChIP-seq; Chemicals; Chromatin; Data; Development; Diabetes Mellitus; Disease; Disease Management; Epilepsy; Excision; Fibroblasts; Functional disorder; Genetic Transcription; Goals; Histones; Human; Human Activities; Individual; Inflammation; Investigation; Knowledge; Laboratories; Language; Lead; Ligands; Malignant Neoplasms; Maps; Masks; Memory; Mitosis; Mitotic; Mitotic Activity; Modeling; Molecular; Mutagenesis; Outcome; Peptides; Pluripotent Stem Cells; Protein Isoforms; Proteins; Reader; Reading; Regenerative Medicine; Regulation; Reporting; Signal Transduction; Tail; Testing; Testis; Virus Replication; chemical binding; chemical function; disorder control; human disease; human pluripotent stem cell; inhibitor; innovation; insight; member; metaplastic cell transformation; molecular modeling; novel; pluripotency; protein protein interaction; stem cells; transcriptome sequencing

Bromodomain extra terminal (BET) proteins are critical readers of the histone acetyl marks, which generally signal active transcription. Human BET proteins include the ubiquitous BRD2, BRD3, and BRD4, as well as the testis-specific BRDt. They are key regulators of mitosis and pluripotency reprogramming among many other cellular and biological processes including transcription, cell cycle, transcriptional memory, and stem cell identity, as well as viral replication and transcription. Bromodomains (BD) in BET proteins are the acetyllysine(KAc)-binding modules capable of binding to acetylated core histones. BET proteins are characterized by two tandem bromodomains (BD-I and BD-II) at their N-termini. Of note, the two tandem BDs within each BET member are much more diverse than the equivalent BDs among BET member proteins. In spite of intense investigation, many fundamental questions about these readers of chromatin acetyl marks remain unsolved. Why are there two BDs within each BET protein? Why are the two tandem BDs of each BET protein more diverse than the equivalent BDs among the BET member proteins? What are the specific acetyl marks of histones each individual BD differentially recognizes? Does each individual BD specifically regulate any biological process? Why do each of human BRD3 and BRD4 has both long and short isoforms? How is the chromatin-reading activity of each BD regulated? As mitotic proteins, how is their mitotic activity tightly controlled? Recent findings in my laboratory have paved an avenue to answering many, if not all, of these fundamental questions. We recently reported that BRD3R, a truncated isoform of human BRD3, uniquely enhances cellular pluripotency reprogramming via regulating mitosis. Our further preliminary data suggest that these BRD3R activities are a result of unmasking of the chromatin-reading activities of the second bromodomain (BD-II) of BRD3/BRD3R due to the absence of the long C-terminal tail. Our additional data suggest that intramolecular masking of the BD-II activities may be a general mechanism among BET proteins. We will study how intramolecular masking/unmasking regulate biological and chemical functions of the two distinct BET bromodomains (Aims 1, 2, and 3). We will also identify the chromatin underpinnings behind the different biological and cellular functions of the two distinct KAc-binding modules of BET proteins (Aim 3). To achieve our goals, we will use up-to-date yet firmly established approaches including the chemically defined reprogramming media, ChIP-seq, ChIP-qPCR, RNA-seq, bioinformatics, histone-tail-peptide pull-down, histone-peptide array, ligand affinity assay, mutagenesis, protein-protein interaction, and others. Dysregulation of the BET KAc-reading activities are implicated in various human diseases such as inflammation, diabetes, and various cancers. The fundamental knowledge gained in this proposed study will provide a scientific basis for the disease control and management for the BET-associated diseases.

溴结构域额外末端（BET）蛋白是组蛋白乙酰标记的关键读取器，通常 信号主动转录。人类BET蛋白包括无处不在的BRD2，BRD3和BRD4，以及 特定于睾丸的BRDT。它们是有丝分裂和多能重新编程的关键调节因子 细胞和生物学过程，包括转录，细胞周期，转录记忆和干细胞 身份以及病毒复制和转录。 BET蛋白中的溴结构域（BD）是 乙酰透析（KAC）结合模块，能够与乙酰化核心组蛋白结合。 BET蛋白是 其特征是在其N末端两个串联溴化域（BD-I和BD-II）。值得注意的是，两个串联BD 在每个BET内部，比BET成员蛋白中的同等BD都要多样化。在 对这些染色质乙酰标记的这些读者的许多基本问题进行了激烈的调查 保持尚未解决。为什么每个BET蛋白质中都有两个BD？为什么每个下注的两个串联BD 蛋白质比BET成员蛋白中的同等BD多样化？什么是特定的乙酰基 组蛋白的标记每个BD都差异化？每个BD是否专门调节 有生物学过程吗？为什么每个人的BRD3和BRD4都有长和短的同工型？怎么样 每个BD调节的染色质阅读活动？作为有丝分裂蛋白，其有丝分裂活性如何紧密 受控？我的实验室的最新发现铺平了一条途径，以回答许多（如果不是全部） 基本问题。我们最近报道说，BRD3R是人类BRD3的截断同工型，独特的 通过调节有丝分裂增强细胞多能重编程。我们进一步的初步数据表明 这些BRD3R活动是揭露第二个染色质阅读活动的结果 由于缺乏长的C末端尾巴，BRD3/BRD3R的溴结构域（BD-II）。我们的其他数据 表明BD-II活性的分子内掩盖可能是BET蛋白的一般机制。 我们将研究分子内掩盖/卸载如何调节两者的生物学和化学功能 独特的BET BET溴化群（目标1、2和3）。我们还将确定背后的染色质基础 BET蛋白的两个不同的KAC结合模块的不同生物学和细胞功能（AIM 3）。到 实现我们的目标，我们将使用最新但坚定确定的方法，包括化学定义的 重编程培养基，芯片序列，芯片QPCR，RNA-seq，生物信息学，组蛋白 - 尾肽下拉， 组蛋白肽阵列，配体亲和力测定，诱变，蛋白质 - 蛋白质相互作用等。失调 赌注读取活动与炎症，糖尿病和 各种癌症。在这项拟议的研究中获得的基本知识将为科学依据 与BET相关疾病的疾病控制和管理。