Mechanisms regulating Evf2 long non-coding RNA transcriptional control

Evf2长非编码RNA转录控制的调控机制

• 批准号: 8297958
• 负责人: Jhumku Dutt Kohtz
• 金额: $ 36.75万
• 依托单位: LURIE CHILDREN'S HOSPITAL OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8297958
• 关键词: Adult; Alleles; Autistic Disorder; Biochemistry; Biological; Brain; Caliber; Cancer Control; Cell Nucleus; Chromatin Remodeling Factor; Complex; DNA; DNA Methylation; DNA Methyltransferase; DNA Modification Methylases; Development; Dimensions; Disease; Dosage Compensation (Genetics); Embryo; Enhancers; Epilepsy; Erinaceidae; Family; Family member; Functional RNA; Gene Dosage; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genome; Genomics; Germ Cells; Grant; Growth; Heterogeneity; Hippocampus (Brain); Human; Imagery; In Vitro; Intergenic DNA; Interneuron function; Interneurons; Laboratory Finding; Lead; Link; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Mental Retardation; Methods; Methylation; Molecular; Molecular Biology; Mono-S; Mus; Nature; Neurons; Neurophysiology - biologic function; Neurosciences; Nuclear; Nuclear Extract; Nuclear RNA; Nucleic Acid Regulatory Sequences; Pattern; Play; Poly A; Prosencephalon; Protein Biochemistry; Publishing; RNA; RNA Biochemistry; RNA Splicing; Recruitment Activity; Regulator Genes; Regulatory Element; Reporting; Role; SMARCA4 gene; Schizophrenia; Techniques; Time; Transcript; Transcription Coactivator; Transcriptional Regulation; Translating; Vertebrates; Work; X Inactivation; base; cell growth; homeodomain; imprint; loss of function; malignant neurologic neoplasms; member; mouse model; mutant; nanometer; nerve stem cell; nervous system disorder; neurodevelopment; neuropsychiatry; news; novel; parental influence; protein complex; research study; structural biology; subventricular zone; transcription factor

DESCRIPTION (provided by applicant): Although a critical role for developmentally regulated transcription factors in patterning and differentiation has been established for some time, the role of long non-coding RNAs (lncRNAs) is only beginning to be understood. My lab discovered novel non-coding RNAs, Evf2 and Evf1, that regulate transcription factors essential for proper GABAergic interneuron development. Our work on Evf2, an lncRNA transcribed as a spliced anti-sense transcript shows that lncRNAs play specific gene regulatory functions. Evf2 is conserved across vertebrates (ultraconserved), and activated by sonic hedgehog in the developing forebrain. We show that Evf2 forms a complex with the DLX2 homeodomain transcription factor and regulates Dlx5/6 enhancer activity. Through analysis of an Evf2 loss-of-function mouse model developed in my lab, we find that Evf2 is not only required for proper transcriptional regulation of Dlx5 and Dlx6, but also for proper GABAergic interneuron development in embryonic brain and adult hippocampal circuitry . While much of my past work has focused on the biological role of Evf2 lncRNA in interneuron development, the precise mechanisms involved in Evf2 long- non-coding RNA-dependent transcriptional regulation remain unknown. The proposed studies will use mouse genetics, RNA and protein biochemistry, and RNA cloud visualization in nuclei of embryonic brain to determine the mechanism of Evf2 lncRNA transcriptional regulation. Some of the techniques are novel, such as the use of RNA cloud visualization in 3 dimensions, mono and bi-allelic analysis, and in vitro DNA-dependent RNA-protein complex assembly; the approach as a whole is novel, as this combination of methods to study lncRNA gene regulation has not been reported. These studies will provide an understanding of a critical aspect of gene regulation with particular relevance to neural development and function, as well as growth control and cancer, where multiple ultraconserved lncRNAs have been identified. PUBLIC HEALTH RELEVANCE: This grant proposes to investigate how the ultraconserved long non-coding RNA Evf2 regulates gene expression in GABAergic interneuron precursors. Since altered GABAergic interneuron function has been linked to epilepsy, autism, schizophrenia, and mental retardation, studies on the normal development of GABAergic interneurons are critical to understanding the molecular basis for multiple neuropsychiatric disorders. In addition, the recent identification of ultraconserved long non-coding RNAs in cancer has made studies of how this class of molecules controls gene regulation relevant to growth control. Ultimately, it is hoped that a better understanding of gene regulation in neuronal subpopulations and cellular growth control will lead to the development of novel therapies for treatment of human neurological diseases and cancer.

描述(申请人提供)：尽管发育调节的转录因子在模式形成和分化中的关键作用已经确定了一段时间，但对长非编码RNA(LncRNAs)的作用才刚刚开始了解。我的实验室发现了新的非编码RNA EVF2和Evf1，它们调控着GABA能中间神经元正常发育所必需的转录因子。我们对EVF2的研究表明，LncRNAs发挥着特定的基因调控功能。EVF2在脊椎动物中是保守的(超低温保存)，并被发育中的前脑中的声波刺猬激活。我们发现EVF2与DLX2同源结构域转录因子形成复合体，并调节Dlx5/6增强子的活性。通过对本实验室建立的EVF2功能丧失小鼠模型的分析，我们发现EVF2不仅对Dlx5和Dlx6的转录调控是必需的，而且对胚胎脑和成年海马区GABA能中间神经元的正常发育也是必需的。虽然我过去的大部分工作都集中在EVF2 lncRNA在神经元间发育中的生物学作用上，但EVF2依赖于长非编码RNA的转录调控的确切机制仍不清楚。拟议的研究将利用小鼠遗传学、RNA和蛋白质生物化学以及胚胎脑核中的RNA云图来确定EVF2 lncRNA转录调控的机制。其中一些技术是新颖的，例如使用三维RNA云可视化，单等位基因和双等位基因分析，以及体外分析 DNA依赖的RNA-蛋白质复合体组装；作为一个整体，这种方法是新的，因为这种方法组合来研究lncRNA基因调控尚未见报道。这些研究将提供对基因调控的一个关键方面的理解，该方面与神经发育和功能以及生长控制和癌症特别相关，在这些方面已经发现了多个超服务的lncRNA。 公共卫生相关性：这项拨款建议调查超常服务的长非编码RNAEVF2如何调节GABA能中间神经元前体的基因表达。由于GABA能中间神经元功能的改变与癫痫、自闭症、精神分裂症和智力低下有关，因此对GABA能中间神经元正常发育的研究对于理解多种神经精神障碍的分子基础至关重要。此外，最近在癌症中发现的超长非编码RNA研究了这类分子如何控制与生长控制相关的基因调控。最终，人们希望更好地了解神经元亚群中的基因调控和细胞生长控制将导致开发治疗人类神经疾病和癌症的新疗法。