Regulation of programmed -1 ribosomal frameshifting by micro-RNAs

micro-RNA 对程序性 -1 核糖体移码的调节

基本信息

批准号：
9278237
负责人：
Jonathan D Dinman
金额：
$ 29.68万
依托单位：
UNIV OF MARYLAND, COLLEGE PARK
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-25 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9278237
关键词：
3-Dimensional AIDS/HIV problem Address Affect Animals Area Base Pairing Basic Science Biological Assay Biological Models CCR5 gene Cell Line Cell physiology Cells Cellular biology Cis-Acting Sequence Codon Nucleotides Collaborations Complex Computer Simulation Computing Methodologies Cytokine Receptors Data Databases Defect Development Developmental Biology Elements Eukaryota Feedback Foundations Gene Expression Genetic Code Genome Global Change HIV-1 Health Human Immune response Immune signaling Immunology In Vitro JAK2 gene Laboratories Link Mathematics Mediating Messenger RNA Methods MicroRNAs Modeling Molecular Molecular Biology Pathway interactions Play Preclinical Drug Evaluation Protein Tyrosine Kinase Proteins Publishing RNA Reading Frames Regulation Reporter Research Resolution Ribosomal Frameshifting Ribosomes Role Sequence Analysis Series Signal Transduction Single Nucleotide Polymorphism Structure Structure-Activity Relationship T-Lymphocyte Technology Telomere Maintenance Terminator Codon Testing Translational Research Untranslated RNA Work Yeasts base biophysical techniques cytokine design drug development frontier innovation mRNA Decay mRNA Transcript Degradation next generation novel premature protein expression response ribosome profiling small molecule therapeutics structural biology therapeutic target unpublished works virology

项目摘要

PROJECT SUMMARY/ABSTRACT Since the genetic code was elucidated in the early 1960's, it has been assumed that mRNAs are always decoded in three base codons, that any deviation from this fundamental rule must be erroneous and thus, deleterious. However, over the past decade, we have shown that a significant fraction of cellular mRNAs harbor cis-acting sequence elements that direct elongating ribosomes to shift reading frame by one base in the 5' (-1) direction. In cellular mRNAs, such Programmed -1 Ribosomal Frameshift (-1 PRF) signals direct ribosomes to premature termination codons where they become substrates for rapid degradation through the Nonsense-Mediated mRNA Decay (NMD) pathway, resulting in decreased expression of the proteins encoded by these mRNAs. Importantly, rates of mRNA degradation are proportional to rates of -1 PRF, a relationship that is conserved in eukaryotes from yeast to humans. Observations from yeast to humans that global changes in -1 PRF rates are deleterious to cellular function suggested that regulation of -1 PRF must be sequence- specific. Recently, we discovered that this is achieved through the interactions between -1 PRF signals and miRNAs. These findings have initiated a completely new avenue of research by establishing -1 PRF on cellular mRNAs, and its regulation by miRNAs as a new, fundamental paradigm in gene expression. This proposal seeks to deepen our understanding of the molecular mechanisms underlying regulation of -1 PRF in human cells. The well-defined Jurkat human T-cell line and a focus on -1 PRF signals embedded in mRNAs encoding cytokine receptors and a critical cytokine-responsive tyrosine kinase provides a model system to address a series of questions ranging from basic molecular and structural biology to regulation and control of the acquired immune response. Aim 1 of this proposal seeks to confirm -1 PRF promoted by sequences identified in the mRNAs encoding IL2Rγ, IL7Rα, and JAK2, characterize the effects of SNPs on -1 PRF, and develop the next generation PRF technology. Aim 2 will identify and validate miRNAs that naturally interact with these -1 PRF signals, and will test an autoregulatory feedback loop model of -1 PRF. Aim 3 is oriented towards characterizing the effects of miRNAs on gene expression and RNA structure. By the end of the proposed studies, we will have 1) deepened our understanding of this new paradigm gene expression control, 2) identified specific miRNAs that are used by T-cells to control their responses to important cytokines, and 3) established new rules describing mRNA/miRNA atomic scale structural interactions. These studies will immediately impact many fields including basic molecular and cell biology, and more applied fields including immunology and HIV/AIDS, and will lay the foundation for the design and discovery of small molecule therapeutics targeted to specific -1 PRF signals.

项目概要/摘要自从 20 世纪 60 年代初阐明遗传密码以来，人们一直认为 mRNA 总是用三个基本密码子解码，任何偏离这一基本规则的行为都一定是错误的，因此，然而，在过去的十年中，我们已经证明很大一部分细胞 mRNA 是有害的。含有顺式作用序列元件，指导延伸核糖体将阅读框移动一个碱基在细胞 mRNA 中，此类程序性 -1 核糖体移码 (-1 PRF) 信号直接向 5' (-1) 方向。核糖体转化为过早终止密码子，并成为快速降解的底物无义介导的 mRNA 衰减 (NMD) 途径，导致编码蛋白的表达减少重要的是，mRNA 降解率与 -1 PRF 的比率成正比，这是一种关系。从酵母到人类的真核生物中保守的从酵母到人类的观察显示全球变化。 -1 PRF 率对细胞功能有害，建议 -1 PRF 的调节必须按顺序进行最近，我们发现这是通过 -1 PRF 信号和之间的相互作用来实现的。这些发现通过在细胞上建立-1 PRF 开启了一条全新的研究途径。 mRNA 及其通过 miRNA 的调控作为基因表达的新的基本范式。旨在加深我们对人类 -1 PRF 调节分子机制的理解明确的 Jurkat 人类 T 细胞系，重点关注编码 mRNA 中的 -1 PRF 信号。细胞因子受体和关键的细胞因子反应性酪氨酸激酶提供了一个模型系统来解决从基本分子和结构生物学到调控的一系列问题该提案的目标 1 旨在确认已识别序列促进的 -1 PRF。在编码 IL2Rγ、IL7Rα 和 JAK2 的 mRNA 中，表征 SNP 对 -1 PRF 的影响，并开发下一代 PRF 技术将识别并验证自然相互作用的 miRNA。这些 -1 PRF 信号，并将测试 -1 PRF 的自动调节反馈环路模型，目标 3。表征 miRNA 对基因表达和 RNA 结构的影响。拟议的研究，我们将1）加深我们对这种新范式基因表达控制的理解， 2) 鉴定出 T 细胞用来控制其对重要细胞因子的反应的特定 miRNA，以及 3) 这些研究将建立描述 mRNA/miRNA 原子尺度结构相互作用的新规则。立即影响许多领域，包括基础分子和细胞生物学，以及更多应用领域，包括免疫学和艾滋病毒/艾滋病，并将为小分子的设计和发现奠定基础针对特定 -1 PRF 信号的治疗。