Tissue biology studies of histone modification, nascent transcription, and post-transcription regulation

组蛋白修饰、新生转录和转录后调控的组织生物学研究

• 批准号: 10746577
• 负责人: JOHN T LIS
• 金额: $ 57.07万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10746577
• 关键词: Address; Adult; Affect; Biological Models; Biological Process; Biology; Cell Lineage; Cell Maintenance; Cell Separation; Cell physiology; Cells; Chromatin; Data; Development; Developmental Gene; Disease; Dissociation; Drug Design; Embryo; Embryonic Development; Enzymes; Epigenetic Process; Epithelium; Future; Gene Expression Regulation; Gene Silencing; Gene Targeting; Genes; Genetic; Genetic Transcription; Genomic approach; Hair; Hair follicle structure; Heterochromatin; Histone H3; Histones; Homeostasis; Impairment; Investigation; Knock-out; Knockout Mice; Knowledge; Maps; Mediating; Messenger RNA; Methodology; Methods; Methyltransferase; Molecular; Morphogenesis; Mus; Natural regeneration; Normal tissue morphology; Nuclear; Pathology; Pattern; Physiological; Polymerase; Population; Post-Transcriptional Regulation; Productivity; RNA; RNA Polymerase II; RNA Probes; RNA Stability; Regulation; Repression; Role; Running; Skin; Stratification; Therapeutic Uses; Time; Tissue Engineering; Tissues; Transcript; Transcription Elongation; Transcription Initiation; Transferase; Work; cell type; conditional knockout; gene repression; genome-wide; histone methylation; histone methyltransferase; histone modification; in vivo; inducible Cre; insight; keratinocyte; liver development; mouse genetics; mouse model; novel; pancreas development; pharmacologic; posttranscriptional; promoter; prototype; recruit; skin organogenesis; tissue mapping; tissue stem cells; tool; transcriptome sequencing; wound healing

Abstract Basic cellular processes essential to mammalian tissue development and adult regeneration, are often controlled by gene expression regulation at the mRNA level. Total mRNA level for each gene depends on two mechanisms: active (or ‘nascent’) transcription and post-transcriptional RNA stability/degradation (e.g. turnover rates). Nascent transcription itself is regulated by multiple steps of recruiting histone-modifying enzymes and opening chromatin, RNA polymerase II (Pol II) initiation and pausing, and the release of Pol II into productive RNA elongation. Diverse mechanisms that regulate total mRNA level may be important in controlling distinct biological processes and pathologies, but this is poorly understood in vivo. Using skin as a model system we will map lineage-specific gene patterns of RNA Pol II activity and nascent transcription in specific cell-types within their natural tissue milieu in the absence of cell isolation. Furthermore, we will investigate changes in these nascent-transcription patterns along an adult tissue stem cell activation and differentiation path, using hair follicle as a lineage prototype. In addition, our work will help dissect relative contributions of active transcription vs post-transcriptional RNA turnover/stability to overall mRNA levels in specific cell types in vivo. Furthermore, we will determine how a known histone repressive mark (H3K9me3), poorly understood in mammalian tissues, acts on nascent transcription and RNA Pol II pausing in vivo. We will uncover for the first time the physiological role of the 3 main H3K9me3 histone methyl transferases in skin development and homeostasis. Finally, we will investigate how H3K9me3 acts on nascent transcription in vivo using our newly developed mouse genetics tools and methodologies. Our work will provide previously unavailable mechanistic insight into gene regulation in mammalian tissue biology using skin as a model system.

摘要 哺乳动物组织发育和成体再生所必需的基本细胞过程是 通常在基因表达水平上受基因表达调控。每种基因的总mRNA水平 基因依赖于两种机制：活跃(或“萌芽”)转录和转录后 RNA稳定性/降解性(例如周转率)。新生的转录本身受 多步骤募集组蛋白修饰酶和开放染色质，RNA聚合酶 II(Pol II)的启动和暂停，以及Pol II的释放进入生产性RNA伸长。多元 调节总信使核糖核酸水平的机制可能在控制不同的生物 过程和病理，但在活体中对此知之甚少。使用外观作为模型系统 我们将定位RNAPOL II活性和新生转录的谱系特异性基因模式 在没有细胞分离的情况下，在其自然组织环境中的特定细胞类型。此外， 我们将沿着成人组织干细胞研究这些新生转录模式的变化。 激活和分化路径，以毛囊为谱系原型。另外，我们的工作 将有助于剖析主动转录与转录后RNA的相对贡献 体内特定细胞类型的整体信使核糖核酸水平的周转/稳定性。此外，我们还将 确定在哺乳动物中知之甚少的已知组蛋白抑制标记(H3K9me3) 组织，作用于体内的新生转录和RNA Pol II暂停。我们将揭开第一个 3种主要H3K9me3组蛋白甲基转移酶在皮肤中的生理作用 发展和动态平衡。最后，我们将调查H3K9me3如何作用于新生 使用我们新开发的小鼠遗传学工具和方法进行体内转录。我们的 这项工作将为哺乳动物的基因调控提供以前无法获得的机械性洞察 以皮肤为模型系统的组织生物学。