Global analysis of hnRNPLL-mediated alternative pre-mRNA splicing in lymphocytes.

淋巴细胞中 hnRNPLL 介导的选择性前 mRNA 剪接的全局分析。

基本信息

批准号：
7713817
负责人：
Shalini Oberdoerffer
金额：
$ 9.72万
依托单位：
IMMUNE DISEASE INSTITUTE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-05-15 至 2010-01-07
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7713817
关键词：
Achievement Affect Alternative Splicing B-Lymphocytes Binding Biological Process Biopolymers CD4 Positive T Lymphocytes CD44 gene Categories Cell Line Cells Collaborations Communities Development Disease Environment Exons Fostering Gene Expression Profile Generations Genes Goals Immune Immune system Institutes Instruction Integral Membrane Protein Knowledge Laboratories Lead Libraries Link Lymphocyte Mature B-Lymphocyte Mediating Mentors Molecular Nuclear PTPRC gene Phase Point Mutation Production Protein Isoforms Protein Tyrosine Phosphatase RNA Interference RNA Splicing Relative (related person)Ribonucleoproteins STAT5A gene Screening procedure Shapes Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Protein Staging T memory cell T-Cell Receptor T-Lymphocyte Technology Trans-Activators Transcript career follow-up human disease in vivo in vivo Model mRNA Precursor novel programs response small hairpin RNA

Achievement Affect Alternative Splicing B-Lymphocytes Binding Biological Process Biopolymers CD4 Positive T Lymphocytes CD44 gene Categories Cell Line Cells Collaborations Communities Development Disease Environment Exons Fostering Gene Expression Profile Generations Genes Goals Immune Immune system Institutes Instruction Integral Membrane Protein Knowledge Laboratories Lead Libraries Link Lymphocyte Mature B-Lymphocyte Mediating Mentors Molecular Nuclear PTPRC gene Phase Point Mutation Production Protein Isoforms Protein Tyrosine Phosphatase RNA Interference RNA Splicing Relative (related person)Ribonucleoproteins STAT5A gene Screening procedure Shapes Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Protein Staging T memory cell T-Cell Receptor T-Lymphocyte Technology Trans-Activators Transcript career follow-up human disease in vivo in vivo Model mRNA Precursor novel programs response small hairpin RNA

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项目摘要

DESCRIPTION (provided by applicant): The generation of distinct protein isoforms as a result of alternative pre-mRNA splicing is particularly prevalent in the immune system. Alternative splicing is reciprocally linked to signal transduction pathways: the majority of transcripts modulated by alternative splicing encode signaling proteins, and trans-acting factors that influence splicing decisions are induced or activated in response to signaling pathways. Alternative splicing of the highly abundant transmembrane protein tyrosine phosphatase, CD45, results in multiple isoforms that are expressed in a stage-specific manner during lymphocyte development. We identified heterogeneous ribonucleoprotein L-like (hnRNPLL) as an inducible, critical regulator of activation- induced CD45 alternative splicing in a splicing factor RNA interference (RNAi) screen. HnRNPLL was necessary and sufficient to promote the transition from CD45RA to CD45RO isoform expression in a T cell line, B cell line and primary T cells. Global analysis of exon level expression revealed that hnRNPLL depletion resulted in alternative exon utilization in a number of additional immune-relevant transcripts, including CD44 and STAT5A. To better understand hnRNPLL activity with respect to CD45 as well as other transcripts, I will 1) explore the determinants of hnRNPLL binding at the molecular level, 2) identify the signaling cascade that culminates in induction of hnRNPLL and 3) modulate hnRNPLL expression in vivo to study its effect on lymphocyte development and function. I will additionally utilize the in vivo models to study the specific activities of the CD45 isoforms. My overall career goal is to spend one to two more year in a mentored phase in the laboratory of Anjana Rao, and then transition into my own laboratory. Anjana's laboratory is the ideal environment to initiate my aims. In addition to a rigorous academic environment, she has access to many cutting edge technologies that will facilitate the achievement of my goals. For instance, we have access to shRNA libraries for screening through the Broad Institute and a Solexa deep sequencer through the Harvard biopolymers facility. Anjana is also very involved in the scientific community and fosters many collaborations. RELEVANCE (See instructions): Given the broad distribution of alternative pre-mRNA splicing, characterization of the link between signaling and alternative splicing is highly significant, especially considering that 10-15% of human diseases are predicted to result from point mutations that affect alternative splicing. Knowledge of the mechanisms that support alternative splicing decisions may ultimately lead to novel therapies for this category of diseases.

描述（由申请人提供）：替代前MRNA剪接的产生不同的蛋白质同工型在免疫系统中尤为普遍。替代剪接与信号转导途径相互链接：大多数由替代剪接编码信号蛋白调节的转录本，以及影响剪接决策的反式作用因子是响应信号通路而诱导或激活的。高度丰富的跨膜蛋白酪氨酸磷酸酶CD45的替代剪接会导致多种同工型在淋巴细胞发育过程中以特异性方式表达。我们确定异质性核糖核蛋白L样（HNRNPLL）是在剪接因子RNA干扰（RNAI）屏幕中激活诱导的CD45替代剪接的诱导，关键调节剂。 HNRNPLL是必要的，足以促进T细胞系，B细胞系和原代T细胞中从CD45RA到CD45RO同工型表达的过渡。外显子水平表达的全球分析表明，HNRNPLL耗竭导致许多其他免疫相关的转录本（包括CD44和STAT5A）的替代外显子利用。为了更好地理解与CD45以及其他转录物相对于HNRNPLL的活性，我将1）探索分子水平上HnRNPLL结合的决定因素，2）确定信号级联，该信号导致最终导致HNRNPLL的诱导和3）调节HNRNPLL在Vivo中的表达来研究其对lympher的效应，以研究其效应lymplys的功能和功能。我还将利用体内模型来研究CD45同工型的特定活性。我的整体职业目标是在Anjana Rao实验室的指导阶段再花一到两年，然后过渡到我自己的实验室。 Anjana的实验室是启动我的目标的理想环境。除了严格的学术环境外，她还可以使用许多尖端技术，这些技术将有助于实现我的目标。例如，我们可以访问通过Broad Institute进行筛查的SHRNA库，并通过哈佛生物聚合物设施进行Solexa Deep Sequencer。 Anjana也非常参与科学界，并培养了许多合作。相关性（请参阅说明）：鉴于替代前MRNA剪接的广泛分布，信号传导与替代剪接之间的联系的表征非常重要，尤其是考虑到10-15％的人类疾病被预测是由于影响替代剪接的点突变而导致的。了解支持替代剪接决策的机制最终可能会导致这种疾病的新疗法。