Regulation of gene expression and genome organization by small RNAs

小RNA调控基因表达和基因组组织

基本信息

批准号：
9330900
负责人：
Carolyn Marie Phillips
金额：
$ 41.25万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2021-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9330900
关键词：
Address Affect Biological Models Caenorhabditis elegans Cell Nucleus Complex Congenital Abnormality Cytology DNA Development Disease Double-Stranded RNA Eukaryota Fire - disasters Gene Expression Gene Expression Regulation Gene Silencing Gene Targeting Genes Genetic Genetic Transcription Genome Genome Stability Goals Human Human Development Infertility Length Light Malignant Neoplasms Messenger RNA MicroRNAs Modification Molecular Molecular Biology Movement Nematoda Nucleotides Organism Pathway interactions Phylogeny Play Positioning Attribute Process Protein Family Proteins Proteomics RNA RNA Interference Recruitment Activity Regulator Genes Research Role Route Set protein Small RNA Transcript Translations Viral Work Yeasts antiviral immunity biochemical tools genome integrity human disease mRNA Stability member programs

Address Affect Biological Models Caenorhabditis elegans Cell Nucleus Complex Congenital Abnormality Cytology DNA Development Disease Double-Stranded RNA Eukaryota Fire - disasters Gene Expression Gene Expression Regulation Gene Silencing Gene Targeting Genes Genetic Genetic Transcription Genome Genome Stability Goals Human Human Development Infertility Length Light Malignant Neoplasms Messenger RNA MicroRNAs Modification Molecular Molecular Biology Movement Nematoda Nucleotides Organism Pathway interactions Phylogeny Play Positioning Attribute Process Protein Family Proteins Proteomics RNA RNA Interference Recruitment Activity Regulator Genes Research Role Route Set protein Small RNA Transcript Translations Viral Work Yeasts antiviral immunity biochemical tools genome integrity human disease mRNA Stability member programs

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项目摘要

Project Summary: Small RNAs regulate gene expression in a wide range of organisms from yeasts to humans and the fundamental mechanisms governing RNA silencing are conserved across most eukaryotes. The term “small RNA” encompasses a diverse set of small regulatory RNAs, ranging from ~18-30 nucleotides in length, all bound by members of the Argonaute protein family. The first small RNAs identified were microRNAs, discovered in C. elegans as regulators of development timing (Ambros and Ruvkun labs). Several years later, the phenomenon of RNA interference (RNAi), gene silencing induced by the introduction of double stranded RNA, was also discovered in C. elegans (Fire and Mello labs). Much work since then has uncovered the roles of a diverse set of proteins that regulate genes by way of small RNAs at the level of transcription, translation, and mRNA stability. By regulating both endogenous and foreign RNAs, small RNAs play a critical role in development, genome stability, and viral defense. The goal of this research program is to investigate the mechanisms by which RNA silencing pathways modulate gene expression and maintain genome integrity. Like Ambros, Ruvkun, Fire, and Mello, we will use the nematode, C. elegans, as a model system to study RNA silencing because worms combine the best of genetic, cytological, molecular, and biochemical tools. There are many fundamental unanswered questions in our understanding of RNA silencing that we plan to address in this proposal. We and others have determined the localization for many factors in the RNA silencing pathway but there has been little to no cytological exploration the RNA and DNA components. Furthermore, the mechanism by which targeted transcripts are recognized and routed into the RNA silencing pathways is unknown. Our first goal is to localize the mRNAs targeted by RNA silencing and to determine the protein requirements for their physical movement from transcription in the nucleus to turnover in the cytoplasmic RNA silencing compartment, the Mutator foci. We will further examine whether RNA silencing affects the subnuclear positioning of the targeted gene loci, to promote mRNA localization and RNA silencing. Despite much work in the field, there are proteins attributed to small RNA pathway for which the mechanism of action is unknown. We also believe there are components of the pathway that have yet to be identified. Our second goal is to use a combination of molecular biology and cytology to examine uncharacterized components of the pathway in the processes of recruitment and retention of RNA in the Mutator foci and in modification of mRNA targets. Our third goal is to identify and characterize new components of the RNA silencing complex using proteomics. This research will shed light on conserved pathways that regulate gene expression and are critical for development, antiviral immunity, and genome organization.

项目摘要：小RNA调节从酵母到各种生物的基因表达在大多数真核生物中，人类和控制RNA沉默的基本机制是保守的。 “小RNA”一词包含一组各种小调节RNA，范围为〜18-30个核苷酸长时间，都受Argonaute蛋白质家族成员的约束。确定的第一个小RNA是 MicroRNA，在秀丽隐杆线虫中被发现为发育时机的调节剂（Ambros和Ruvkun Labs）。一些多年后，RNA干扰现象（RNAi），通过引入双重诱导的基因沉默在秀丽隐杆线虫（Fire and Mello Labs）中也发现了滞留的RNA。从那时起，很多工作已经发现一组各种蛋白质的作用，这些蛋白质在转录水平上通过小RNA调节基因的作用，翻译和mRNA稳定性。通过控制内源性和外源RNA，小RNA起着关键在发展，基因组稳定和病毒防御中的作用。该研究计划的目的是研究RNA沉默途径的机制调节基因表达并维持基因组完整性。像Ambros，Ruvkun，Fire和Mello一样，我们将使用线虫，秀丽隐杆线虫作为研究RNA沉默的模型系统，因为蠕虫结合了最好的遗传，细胞学，分子和生化工具。在我们计划在此提案中解决的RNA沉默。我们和其他人已经确定了RNA沉默途径中许多因素的本地化，但是 RNA和DNA成分几乎没有细胞学探索。此外，该机制识别哪些靶向转录物并将其路由到RNA沉默途径中是未知的。我们的第一个目标是定位由RNA沉默靶向的mRNA，并确定其蛋白质要求从核中转录到细胞质RNA沉默中的营业额的物理运动隔室，突变灶。我们将进一步检查RNA沉默是否影响下核靶向基因定位的定位，以促进mRNA定位和RNA沉默。尽管该领域有很多工作，但仍有蛋白质归因于小的RNA途径作用机理尚不清楚。我们还认为，途径的组成部分尚未确定。我们的第二个目标是使用分子生物学和细胞学的组合来检查在募集和保留RNA的过程中，该途径的未表征成分突变灶和mRNA靶标的修饰。我们的第三个目标是识别和描述新的目标使用蛋白质组学的RNA沉默复合物的成分。这项研究将阐明保守调节基因表达并且对于发育，抗病毒免疫和基因组至关重要的途径组织。