Novel Regulators of T Cell mRNA Decay

T 细胞 mRNA 衰变的新型调节剂

• 批准号: 7792496
• 负责人: Paul R Bohjanen
• 金额: $ 32.9万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7792496
• 关键词: 3' Untranslated Regions; Affinity Chromatography; Apoptosis; Applications Grants; Autoimmune Diseases; Autoimmunity; Be++ element; Beryllium; Binding; Binding Proteins; Biochemical; Biological; Biological Assay; Cell Cycle Progression; Cells; Communicable Diseases; Computing Methodologies; Conserved Sequence; Cytokine Gene; Databases; Development; Disease; Elements; Embryo; Equilibrium; Event; FOS gene; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Grant; Growth; Hela Cells; Homologous Protein; Human; Immunodeficiency and Cancer; Indium; Laboratories; Lead; Life; Malignant Neoplasms; Measures; Mediating; Messenger RNA; Methods; Microarray Analysis; Molecular; Pathway interactions; Patients; Pharmaceutical Preparations; Play; Post-Transcriptional Regulation; Process; Production; Protein Binding; Proteins; Proto-Oncogenes; RNA; RNA Decay; RNA Sequences; RNA-Binding Proteins; Regulation; Regulator Genes; Regulatory Element; Reporter; Rest; Ribonucleic Acid Regulatory Sequences; Role; Sequence Analysis; System; T cell regulation; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Testing; Time; Transcript; Transcription factor genes; Transcriptional Regulation; Translations; Xenopus; base; c-myc Genes; human disease; insight; mRNA Decay; mRNA Transcript Degradation; novel; receptor; research study

DESCRIPTION (provided by applicant): The broad long-term goal of this project is to understand the role of mRNA degradation in regulating T lymphocyte gene expression. Steady state mRNA levels are determined by the balance between transcription and degradation. This proposal is based on the hypothesis that specific RNA regulatory sequences and specific trans-acting proteins that recognize these sequences function to regulate gene expression at the level of mRNA decay. We recently used microarray technology to measure mRNA decay rates of approximately 6,000 transcripts expressed in T cells under conditions of rest or activation, and we identified hundreds of transcripts that were regulated at the level of mRNA decay. We used computational methods to search for conserved sequences found in transcripts that were co-regulated at the level of mRNA decay and found that approximately 100 short-lived transcripts expressed in T cells contained the conserved 11-mer RNA sequence UGUUUGUUUGU in their 3' untranslated region. We have termed this sequence the GU-rich element (GRE) and have shown using a reporter system that this RNA element mediates mRNA decay in HeLa cells. In preliminary experiments, we have also shown that the CUG-binding protein 1 binds specifically to the GRE and mediates GRE-dependent mRNA decay. The goals of this grant are to characterize these novel regulators and to identify additional novel regulators of mRNA decay. Specific aim 1 is to characterize the role of the GU-rich element (GRE) and the CUG-binding protein 1 (CUGBP1) in regulating T cell mRNA decay. Specific aim 2 is to characterize the role of novel conserved RNA sequence elements in regulating T cell mRNA decay, and specific aim 3 is to characterize the role of novel RNA-binding proteins in regulating T cell mRNA decay. We will use biochemical and molecular biological methods to test the function of novel regulatory elements and RNA-binding proteins in the regulation of T cell mRNA decay. These experiments will provide insight into the mechanisms by which mRNA decay regulates the expression of important growth regulatory genes, including cytokine genes and proto-oncogenes. Understanding the mechanisms by which mRNA decay regulates the expression of these genes will provide insight into human disease states such as autoimmunity, immunodeficiency, cancer, and infectious diseases and will lead to the development of new anti-cancer and immunomodulatory drugs that act by targeting specific mRNA decay pathways. In order for human cells to develop and function normally, genes need to be turned on at precise times and they need to be turned off at precise times. An important method that cells use to turn genes off is to eliminate messenger RNA, which then stops the production of proteins. The elimination of messenger RNA occurs in the cell through a process known as messenger RNA decay. The goal of the experiments proposed in this grant application is to define the proteins that are responsible for mRNA decay and to understand how the process of mRNA decay is regulated. Many human diseases such as cancer or autoimmune diseases occur because genes are turned on but then are not turned off in a normal manner. Understanding how genes are turned off will lead to the development of new drugs that will be used to turn off specific genes in patients with diseases such as cancer or autoimmunity.

描述（由申请人提供）：该项目的广泛长期目标是了解mRNA降解在调节T淋巴细胞基因表达中的作用。稳态mRNA水平取决于转录和降解之间的平衡。该建议基于以下假设：识别这些序列在mRNA衰减水平下调节基因表达的特定RNA调节序列和特定的反式蛋白质。我们最近使用微阵列技术来测量在休息或激活条件下在T细胞中表达的大约6,000次转录本的mRNA衰减速率，我们确定了数百个在mRNA衰减水平下受调节的转录本。我们使用计算方法来搜索在mRNA衰变水平上共同调节的转录本中发现的保守序列，并发现在T细胞中表达的大约100个短寿命转录本包含保守的11-mer RNA序列Uguuuuguuguugu在其3'未经翻译区域中。我们已经将此序列称为富含GU的元素（GRE），并使用报告基因系统表明该RNA元素介导HeLa细胞中的mRNA衰变。在初步实验中，我们还表明，CUG结合蛋白1特异性与GRE结合并介导GRE依赖性mRNA衰变。这项赠款的目标是表征这些新型调节器，并确定mRNA衰变的其他新型调节剂。具体目的1是表征富含GU的元素（GRE）和CUG结合蛋白1（CUGBP1）在调节T细胞mRNA衰变中的作用。具体目的2是表征新型保守的RNA序列元件在调节T细胞mRNA衰减中的作用，而特定的目标3是表征新型RNA结合蛋白在调节T细胞mRNA衰变中的作用。我们将使用生化和分子生物学方法来测试新型调节元件和RNA结合蛋白在T细胞mRNA衰变调节中的功能。这些实验将提供有关mRNA衰变调节重要生长调节基因（包括细胞因子基因和原始癌基因）表达的机制的洞察力。了解mRNA衰变调节这些基因表达的机制将提供对人类疾病状态的见解，例如自身免疫性，免疫缺陷，癌症和感染性疾病，并将导致新的抗癌和免疫调节药物的发展，这些药物通过针对特定的特定mRNA衰变途径来起作用。为了使人类细胞正常发育和功能，需要精确打开基因，并且需要在精确的时间关闭它们。细胞用来关闭基因的一种重要方法是消除信使RNA，然后阻止蛋白质的产生。通过称为Messenger RNA衰减的过程，消除了信使RNA。本赠款应用中提出的实验的目的是定义负责mRNA衰变的蛋白质，并了解如何调节mRNA衰变过程。由于基因被打开，但不会以正常方式关闭，因此发生了许多人类疾病，例如癌症或自身免疫性疾病。了解如何关闭基因将导致新药的发展，这些药物将用于关闭癌症或自身免疫性疾病患者的特定基因。