Novel Regulators of T Cell mRNA Decay

T 细胞 mRNA 衰变的新型调节剂

• 批准号: 8104636
• 负责人: Paul R Bohjanen
• 金额: $ 1.49万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-15 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8104636
• 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 水平由转录和降解之间的平衡决定。该提议基于这样的假设：特定的RNA调控序列和识别这些序列的特定反式作用蛋白在mRNA衰变水平上发挥调节基因表达的作用。我们最近使用微阵列技术测量了 T 细胞在静止或激活条件下表达的约 6,000 个转录本的 mRNA 衰减率，并鉴定了数百个在 mRNA 衰减水平上受到调节的转录本。我们使用计算方法来搜索在 mRNA 衰减水平上共同调控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 个短寿命转录本在其 3' 非翻译中包含保守的 11 聚体 RNA 序列 UGUUUGUUUGU地区。我们将该序列称为富含 GU 元件 (GRE)，并使用报告系统证明该 RNA 元件介导 HeLa 细胞中的 mRNA 衰减。在初步实验中，我们还表明 CUG 结合蛋白 1 特异性结合 GRE 并介导 GRE 依赖性 mRNA 衰减。这笔资助的目标是描述这些新型调节因子的特征，并确定其他 mRNA 衰减的新型调节因子。具体目标 1 是表征富含 GU 的元件 (GRE) 和 CUG 结合蛋白 1 (CUGBP1) 在调节 T 细胞 mRNA 衰减中的作用。具体目标 2 是表征新型保守 RNA 序列元件在调节 T 细胞 mRNA 衰减中的作用，具体目标 3 是表征新型 RNA 结合蛋白在调节 T 细胞 mRNA 衰减中的作用。我们将利用生化和分子生物学方法来测试新型调控元件和RNA结合蛋白在T细胞mRNA衰减调控中的功能。这些实验将深入了解 mRNA 衰减调节重要生长调节基因（包括细胞因子基因和原癌基因）表达的机制。了解 mRNA 衰变调节这些基因表达的机制将有助于深入了解自身免疫、免疫缺陷、癌症和传染病等人类疾病状态，并将导致开发针对特定基因发挥作用的新型抗癌和免疫调节药物。 mRNA 衰变途径。为了使人体细胞正常发育和发挥功能，基因需要在精确的时间打开和关闭。细胞关闭基因的一个重要方法是消除信使 RNA，从而停止蛋白质的产生。细胞中信使 RNA 的消除通过称为信使 RNA 衰减的过程发生。本次资助申请中提出的实验目标是确定导致 mRNA 衰变的蛋白质，并了解 mRNA 衰变过程是如何受到调节的。许多人类疾病，例如癌症或自身免疫性疾病，都是因为基因被打开但随后没有以正常方式关闭而发生。了解基因如何关闭将有助于开发新药物，用于关闭患有癌症或自身免疫等疾病的患者的特定基因。