The Positive Roles of Lysine Deacetylase Complexes in Regulating Transcriptional Dynamics (Diversity Supplement)

赖氨酸脱乙酰酶复合物在调节转录动力学中的积极作用（多样性补充）

• 批准号: 10612509
• 负责人: Catharine Lynn Smith
• 金额: $ 1.63万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10612509
• 关键词: Acetylation; Acetyltransferase; Biological Models; Cell physiology; Cells; Clinical; Complex; DNA Polymerase II; Deacetylase; Disease; Drug Targeting; Enhancers; Genes; Genetic Transcription; Glucocorticoid Receptor; Glucocorticoids; Goals; Histone Acetylation; Histone Deacetylase; Human Pathology; Investigation; Kinetics; Knowledge; Lysine; Measures; Mediating; Modeling; Pattern; Process; Public Health; Publishing; RNA; RNA Polymerase II; Research; Role; Signal Transduction; Testing; Transcription Coactivator; Transcription Initiation; Transcription Initiation Site; Transcriptional Activation; Transcriptional Regulation; Work; career; epigenome; gene repression; genetic corepressor; genome-wide; glucocorticoid receptor alpha; graduate student; inhibitor; nanobodies; novel; parent grant; promoter; recruit; skills; therapeutic development

PROJECT SUMMARY Whereas traditional models of transcription cast lysine acetyltransferases (KATs, also known as HATs) as tran- scriptional coactivators and lysine deacetylases (KDACs, also known as HDACs) as corepressors, an abun- dance of evidence demonstrates that KDACs can facilitate transcription in a gene-dependent fashion. However, the mechanisms underlying their transcription-promoting functions are poorly understood. The long-term goal of our work is to define mechanisms by which KDACs, KATs, and acetylation regulate signaling-modulated tran- scription. The proposed study utilizes glucocorticoid signaling as a model system. Published studies from the lab demonstrate that KDACs are required not only for glucocorticoid-mediated transcriptional repression but also for transcriptional activation of target genes. Preliminary studies show that KDAC1 is required for GR-activated transcription by multiple mechanisms, depending on the target gene, acting either upstream or downstream of RNA polymerase II (RNA Pol II) recruitment to the transcription start site (TSS). The next logical step is to identify the KDAC1 complexes involved and investigate their roles in glucocorticoid receptor (GR)-activated transcription. The objective of the parent grant (R01GM139829) is to define the roles of KDACs in the dynamics of the tran- scriptional cycle at glucocorticoid receptor (GR)-activated genes. The objective of the research plan described in this diversity supplement is to investigate the impact of KDAC activity on the dynamics of RNA polymerase II association with GR-activated genes. This hypothesis will be tested experimentally through three specific aims. In the first specific aim, the impact of Class I KDACs on the kinetics of transcriptional bursting at GR-activated genes will be measured to identify GR-activated genes that are regulated by KDAC activity upstream or down- stream of transcriptional initiation, working from the hypothesis that the mechanism by which KDACs facilitate transcription is gene-specific. In the second specific aim, the impact of KDAC activity on RNA pol II distribution and the pattern of nascent transcription at enhancers, promoters, and gene bodies of GR-activated genes will be determined genome-wide. The working hypothesis is that KDAC activity regulates transcription and RNA pol II dynamics at distinct gene regions in a gene-specific fashion. In the third specific aim the dynamics of histone acetylation and RNA pol II will be investigated at a GR-activated gene at which transcription can be visualized in live cells. This will be performed using nanobodies to detect histone acetylation and RNA pol II in live cells. This work will generate novel mechanistic knowledge of the transcriptional functions of KDACs that is relevant to basic understanding of cellular processes as well as treatment of disease through modulation of the epige- nome. Importantly, in the process of doing this research the graduate student who will be supported by the supplement will gain skills critical to a successful career in the biomedical workforce.

项目摘要 而传统的转录模型将赖氨酸乙酰转移酶（KAT，也称为HAT）作为转基因生物。 作为辅阻遏物的转录辅激活因子和赖氨酸脱乙酰酶（KDAC，也称为HDAC）， 大量证据表明，KDAC可以以基因依赖的方式促进转录。然而，在这方面， 其转录促进功能的机制知之甚少。的长期目标 我们的工作是确定KDAC，KAT和乙酰化调节信号调节的转录的机制， 笔迹。该研究利用糖皮质激素信号作为模型系统。实验室发表的研究 表明KDAC不仅是糖皮质激素介导的转录抑制所必需的，而且也是 靶基因的转录激活。初步研究表明，KDAC 1是GR激活所必需的。 通过多种机制转录，这取决于靶基因，作用于上游或下游。 RNA聚合酶II（RNA Pol II）募集至转录起始位点（TSS）。下一个合乎逻辑的步骤是确定 KDAC 1复合物参与糖皮质激素受体（GR）激活的转录，并研究其作用。 父母补助金（R01GM139829）的目的是定义KDAC在跨文化交流动态中的作用， 糖皮质激素受体（GR）激活基因的转录周期。研究计划的目标描述 在这个多样性补充是调查的影响，KDAC活动的动态RNA聚合酶II 与GR激活基因的关联。这一假设将通过三个具体目标进行实验检验。 在第一个具体目标中，研究了I类KDAC对GR激活的转录爆发动力学的影响。 将测量基因以鉴定由KDAC活性上游或下游调节的GR激活基因。 转录起始流，从KDAC促进转录的机制这一假设出发， 转录是基因特异性的。在第二个具体目标中，KDAC活性对RNA聚合酶II分布的影响 GR激活基因的增强子、启动子和基因体处的新生转录模式将 全基因组决定。工作假设是KDAC活性调节转录和RNA聚合。 II动力学在不同的基因区域的基因特异性的方式。在第三个具体目标中， 乙酰化和RNA聚合酶II将在GR激活的基因中进行研究，在该基因处可以观察到转录 在活细胞中。这将使用纳米抗体来检测活细胞中的组蛋白乙酰化和RNA pol II。 这项工作将产生新的机制知识的转录功能的KDAC是相关的 对细胞过程的基本理解，以及通过调节表皮来治疗疾病， 诺姆。重要的是，在做这项研究的过程中，研究生谁将得到支持， 补充将获得关键的技能，在生物医学劳动力的成功职业生涯。