TAFopathies Result from Derangements in Transcriptional Control of Metabolism

TAF 病是由代谢转录控制紊乱引起的

• 批准号: 10314285
• 负责人: Jamison Leid
• 金额: $ 3.2万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10314285
• 关键词: TAFopathies; Result; Derangements; Transcriptional; Control

Project Summary Intellectual disability is a disabling neurodevelopmental disorder that affects 2-3% of the general population. Often times, these intellectual disabilities are accompanied by additional developmental abnormalities. These syndromic forms of intellectual disability frequently have a genetic basis. Recent studies have identified a subset of intellectual disability patients with pathogenic variants in key components of the TATA-binding protein associated factors (TAFs). These patients also have craniofacial defects and congenital heart disease. This syndrome has been termed as a TAFopathy and includes mutations in TATA binding protein (TBP), TAF1, TAF2, and TAF6. Through a forward genetic screen, we have recovered a lethal nonsense mutation in TAF5. Mutant embryos have craniofacial hypoplasia, ventricular hypoplasia, and heart failure at 96 hours post fertilization. CRISPR/Cas9 mediated gene editing revealed that this phenotype was recapitulated in TAF1 KO and TAF5 KO embryos. TAF5 KOs show significant metabolic dysfunction that may precede the anatomical defects observed at 96 hours post fertilization. Together, these findings support a regulatory role for TAFs in coordinating transcriptional activation of metabolic programming during embryogenesis. The TAFs form the general transcription factor TFIID, which recruits RNA Polymerase II to form the preinitation complex at sites of transcription. Studies have shown that individual TAFs are not necessary for general transcription. Other studies have identified functional domains of TAF1 that activate transcription through interactions with TBP and enhancer elements that are sufficient to recruit RNA Polymerase II to sites of transcription. In conjunction with our data showing metabolic dysfunction, we hypothesize that TAFs regulate metabolic programs by linking enhancers and promoters of metabolic genes through TFIID assembly. Our current efforts are focused towards completing our phenotypic characterization of TAF1 KOs and TAF5 KOs with respect to craniofacial- and neuro-development. We plan to use innovative and cutting-edge techniques, including x-ray microscopy, RNA- Scope in situ hybridization, mitochondrial respiration assays, ChIP-seq, and Hi-C to achieve this goal. The data obtained will help us elucidate basic disease mechanisms of TAFopathy. These findings would also indicate a secondary function for general transcription factors to regulate metabolic programming during embryogenesis. .

项目概要 智力障碍是一种致残性神经发育障碍，影响 2-3% 的总人口。 很多时候，这些智力障碍还伴随着额外的发育异常。这些 智力障碍的综合症形式通常有遗传基础。最近的研究已经确定了一个 TATA 结合蛋白关键成分存在致病性变异的智力障碍患者亚群 相关因素（TAF）。这些患者还患有颅面部缺陷和先天性心脏病。这 该综合征被称为 TAF 病，包括 TATA 结合蛋白 (TBP)、TAF1、 TAF2 和 TAF6。通过正向遗传筛选，我们在 TAF5 中发现了致命的无义突变。 突变胚胎在96小时后出现颅面发育不全、心室发育不全和心力衰竭 施肥。 CRISPR/Cas9 介导的基因编辑揭示了这种表型在 TAF1 KO 中得到了重现 和TAF5 KO胚胎。 TAF5 KOs 显示出明显的代谢功能障碍，可能先于解剖学 受精后96小时观察到的缺陷。总之，这些发现支持 TAF 在以下方面的监管作用： 协调胚胎发生过程中代谢程序的转录激活。 TAF 组成 一般转录因子 TFIID，招募 RNA 聚合酶 II 在以下位点形成预起始复合物 转录。研究表明，单个 TAF 对于一般转录来说并不是必需的。其他 研究已确定 TAF1 的功能域通过与 TBP 相互作用来激活转录 足以将 RNA 聚合酶 II 招募到转录位点的增强子元件。结合 我们的数据显示代谢功能障碍，我们假设 TAF 通过链接来调节代谢程序 通过 TFIID 组装代谢基因的增强子和启动子。我们目前的努力重点是 完成我们对 TAF1 KO 和 TAF5 KO 在颅面和面部的表型表征 神经发育。我们计划使用创新和尖端技术，包括 X 射线显微镜、RNA- 通过原位杂交、线粒体呼吸测定、ChIP-seq 和 Hi-C 来实现这一目标。数据 所获得的结果将有助于我们阐明TAF病的基本发病机制。这些发现也表明 一般转录因子的次要功能是调节胚胎发生过程中的代谢程序。 。