Regulation of programmed -1 ribosomal frameshifting by micro-RNAs

micro-RNA 对程序性 -1 核糖体移码的调节

• 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
• 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protein expression; response; ribosome profiling; small molecule therapeutics; structural biology; therapeutic target; unpublished works; virology

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.

项目摘要/摘要 由于遗传密码在1960年代初期被阐明，因此人们认为mRNA始终是 在三个基本密码子中解码，任何偏离此基本规则都必须是错误的，因此 有害。但是，在过去的十年中，我们已经表明了很大一部分细胞mRNA 携带顺式作用序列元件，将核糖体伸长以将阅读框移到一个基础上 5'（-1）方向。在细胞mRNA中，此类编程的-1核糖体移状（-1 PRF）信号直接 核糖体到过早终止密码子，它们成为通过 废话介导的mRNA衰变（NMD）途径，导致蛋白质的表达降低 通过这些mRNA。重要的是，mRNA降解的速率与-1 prf的速率成正比，关系 从酵母到人类的真核生物中，这是保守的。从酵母到人类的观察，全球变化 在-1中，将PRF速率删除到细胞功能表明-1的调节必须是序列 - 具体的。最近，我们发现这是通过-1 prf信号与 mirnas。这些发现通过建立-1 prf的细胞来启动了全新的研究途径 mRNA及其由miRNA作为基因表达的新的基本范式的调节。这个建议 旨在加深我们对人类-1 PRF基本机制的分子机制的理解 细胞。定义明确的Jurkat人类T细胞系列，重点是嵌入MRNA编码中的-1 PRF信号 细胞因子受体和关键的细胞因子响应性酪氨酸激酶提供了一种模型系统来解决 一系列问题，从基本分子和结构生物学到调节和控制 获得的免疫响应。本提案的目标1旨在确认-1 prf通过确定的序列促进 在编码IL2Rγ，IL7Rα和JAK2的mRNA中，表征了SNP对-1 PRF的影响，并发展 下一代PRF技术。 AIM 2将识别和验证自然相互作用的miRNA 这些-1个PRF信号，并将测试-1 PRF的自动调节反馈循环模型。 AIM 3是定向的 旨在表征miRNA对基因表达和RNA结构的影响。到结束 拟议的研究，我们将有1）加深对这种新范式基因表达控制的理解， 2）鉴定出T细胞用于控制其对重要细胞因子的反应的特定miRNA，3） 建立了描述mRNA/miRNA原子尺度结构相互作用的新规则。这些研究会 立即影响许多领域，包括基本分子和细胞生物学，以及更多应用领域 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 治疗针对特定-1 PRF信号。