An inquiry into the function of Ronin in embryogenesis and pluripotent stem cells

Ronin在胚胎发生和多能干细胞中的功能探究

• 批准号: 9897561
• 负责人: Thomas P. Zwaka
• 金额: $ 46.61万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-11 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9897561
• 关键词: Apoptosis; Binding; Binding Sites; Cancer Biology; Cell Nucleus; Cells; Chromatin Loop; Chromosome Mapping; Communication; Complex; Congenital Abnormality; DNA Binding; DNA Binding Domain; DNA Transposons; DNA-Binding Proteins; Data; Defect; Development; Developmental Gene; Dimerization; Disease; Distant; Elements; Embryonic Development; Enhancers; Event; Evolution; Family; Family member; Gene Cluster; Gene Expression; Gene Expression Regulation; Gene Family; Genes; Genetic Enhancer Element; Genetic Transcription; Genome; Heart; Heart Diseases; Human; In Vitro; Link; Maintenance; Mediating; Medicine; Molecular; Mustard; Nematoda; Neurologic; Nuclear Protein; Organelles; Phosphorus; Pluripotent Stem Cells; Process; Promoter Regions; Proteins; Recording of previous events; Regulator Genes; Reporting; Resolution; Stem Cell Development; Testing; Traction; Transactivation; Transcriptional Regulation; Ursidae Family; Zinc Fingers; cancer type; combinatorial; embryonic stem cell; gene interaction; in vivo; insight; member; organ growth; pluripotency; promoter; recruit; research study; stem cell biology; stem cells; transcription factor

PROJECT SUMMARY A long-held view is that gene regulatory complexity in stem cells and development is achieved through the judicious organization of millions of enhancer elements. A recently identified new class of DNA binding fac- tors called THAPs (Thanatos-associated domain-containing apoptosis-associated proteins) may provide new insights into gene regulation in stem cells. One key member of this family, Ronin (Thap11), was discovered by us in embryonic stem cells. Ronin is expressed during the earliest stages of embryonic development and is essential for the maintenance of stem cell pluripotency, both in vitro and in vivo. Since Ronin lacks appreciable transac- tivation activity but functions as a DNA-binding nuclear protein with strong protein dimerization activity, we considered that it may not act in the classical manner of a transcriptional regulator (e.g., having an acidic activa- tion domain) but instead might augment gene expression by facilitating communication among Ronin binding sites through DNA looping. This working hypothesis gained traction when we found that most (83%) of the looping events mediated by Ronin involve promoter-to-promoter links rather than the enhancer-to-promoter interactions seen with typical transcription factors. In this research study we will (i) establish the effects of Ro- nin-mediated promoter tethering on transcription and gene regulation in stem cells; (ii) determine the influence of promoter tethering on developmental transitions especially from the naïve to primed pluripotent state; and (iii) ask if other THAPs participate in development specific gene clustering as well.

项目摘要 长期以来的观点是，干细胞中的基因调节复杂性，并通过 明智地组织了数百万个增强子元素。最近确定的新类DNA结合FAC- 称为THAPS（与Thanatos相关结构域相关蛋白的TOR）可能会提供新的 对干细胞中基因调节的见解。我们发现了这个家庭的一个关键成员Ronin（THAP11）。 在胚胎干细胞中。罗宁在胚胎发育的最早阶段表达，这是必不可少的 为了维持干细胞多能在体外和体内。由于罗宁缺乏明显的转换 TIVATIOT活性，但起着具有强蛋白二聚作用的DNA结合核蛋白的作用，我们 认为它可能不会以转录调节剂的经典方式起作用（例如，具有酸性活化 tion域），但可以通过支持Ronin结合之间的交流来增强基因表达 通过DNA循环的位置。当我们发现大多数（83％） 由Ronin介导的循环事件涉及促进者到启动器链接，而不是增强器到促销。 与典型转录因子看到的相互作用。在这项研究中，我们将（i）确定RO-的影响 NIN介导的启动子在干细胞中的转录和基因调节方面的束缚； （ii）确定影响 促进者在发育过渡上的束缚，尤其是从幼稚到底漆的多能状态；和 （iii）询问其他THAP是否也参与特定的基因聚类。