Cytokine Gene Regulation by Modification of Arginine Residues

通过精氨酸残基修饰进行细胞因子基因调控

基本信息

批准号：
8075289
负责人：
KERRI A MOWEN
金额：
$ 1.1万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-06-01 至 2010-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8075289
关键词：
Allergic Amino Acid Sequence Amino Acids Antibodies Arginine Arginine deiminase Autoimmune Diseases Autoimmunity Binding Biological Assay Cell physiology Cells Citrulline Cytokine Gene Data Ectopic Expression Event Exons Gene Expression Regulation Gene Targeting Glutamine Histones Human Hydrolysis Immune Immune response Immunologic Receptors In Vitro Inflammatory Interleukin-4 Ligation Link Lymphocyte Mass Spectrum Analysis Mediating Methylation Modification Molecular Mono-S Mus Mutate Nuclear Protein Nuclear Proteins Pattern Phenotype Positioning Attribute Post-Translational Protein Processing Predisposition Production Protein-arginine deiminase Proteins Receptor Signaling Regulation Reporter Genes Research Personnel Rheumatoid Arthritis Role Signal Pathway Site T-Cell Receptor T-Lymphocyte TRAF2 gene Testing Th2 Cells arginine methyltransferase base cell type cofactor cytokine genetic regulatory protein in vivo methyl group mutant novel overexpression prevent programs response tool

项目摘要

Cytokine production by the Th1 and Th2 subsets have been associated with susceptiblity to infectious, allergic, and autoimmune diseases. Understanding the molecular events which control lineage-specific cytokine expression would provide useful tools to modulate the Th1/Th2 response. IL- 4 production by Th2 cells is regulated by NFAT and the NFAT cofactor, NIP45. We have shown that arginine methylation of NIP45 facilitates its interaction with NFAT and augments IL-4transcription. Our data positioned the arginine methyltransferase PRMT1 downstream of the T cell receptor, suggesting that arginine methylation may be an important modification in immune receptor signaling pathways. Arginine methylation is countered by the actions of peptidylarginine deiminase 4 (PAD4). The five PAD enzymes convert arginine residues within proteins into the atypical amino acid citrulline. PAD4 is expressed mainly in lymphocytes. We have found that NIP45 methylation is negatively regulated by PAD4 via citrullination of arginine residues, which prevents the ability of NIP45 to be methylated by PRMT1. PAD4 expression dramatically reduces NIP45-induced IL-4promoter activity. Therefore, we hypothesize that through actions on NIP45 and other regulatory proteins that PAD4 controls Th cell cytokine expression.The specific aimsare: 1) Determine the mechanism by which PAD4 antagonizes NIP45-mediatedinduction of Th cell cytokine production. We will (i) determine the modified arginine residues in NIP45 by mass spectrometry, (ii) determine how PAD4 influences NIP45 activity by studying the effects of PAD4 on the NIP45/NFAT interaction, (iii) determine whether citrullination within NIP45 serves a function other than preventing methylation. 2) Investigate the regulation of PAD4 activity. We will: (i) determine the PAD4 expression pattern in Th cells, (ii) determine whether PAD4 activity is regulated in Th cells, (iii) determine whether PAD4, itself, is regulated by arginine methylation. 3) Analyzethe phenotype of PAD4 deficient mice. We will create mice in which exons 7-13 of PAD4 are flanked by loxP sites so that we can ablate PAD4 expression in the T lineage using CD4-Cre Tg mice. These mice will allow us to determine the role of PAD4 in Th cell function.

TH1和TH2子集的细胞因子产生与敏感性有关感染性，过敏性和自身免疫性疾病。了解控制的分子事件谱系特异性细胞因子表达将提供有用的工具来调节TH1/TH2响应。 il- 4 Th2细胞的产生由NFAT和NFAT辅因子NIP45调节。我们已经表明 NIP45的精氨酸甲基化有助于其与NFAT的相互作用，并增强IL-4转录。我们的数据将T细胞受体下游的精氨酸甲基转移酶PRMT1定位表明精氨酸甲基化可能是免疫受体信号传导的重要修饰途径。精氨酸甲基化被肽基金蛋白脱节酶4（PAD4）的作用抵消。五种垫酶将蛋白质中的精氨酸残基转化为非典型氨基酸瓜氨酸。 PAD4主要在淋巴细胞中表达。我们发现NIP45甲基化是负面的通过精氨酸残基的柠檬酸来调节PAD4，这阻止了Nip45的能力由PRMT1甲基化。 PAD4表达大大降低了NIP45诱导的IL-4促进性活性。因此，我们假设通过对NIP45和其他调节蛋白的作用来假设 PAD4控制细胞因子的表达。特定的目标： 1）确定PAD4拮抗Nip45介导的TH细胞的机制细胞因子产生。我们将（i）通过质量确定NIP45中修饰的精氨酸残基光谱法，（ii）通过研究PAD4的影响来确定PAD4如何影响NIP45活性在NIP45/NFAT相互作用上，（iii）确定NIP45中的柠檬酸是否为a 除了防止甲基化以外的功能。 2）研究PAD4活性的调节。我们将：（i）确定PAD4表达模式在TH细胞中，（ii）确定PAD4活性是否在TH细胞中受到调节，（iii）确定是否是否 PAD4本身受精氨酸甲基化调节。 3）分析PAD4缺乏小鼠的表型。我们将创建老鼠，其中外显子7-13 PAD4是LOXP位点的两侧，因此我们可以使用PAD4的表达 CD4-CRE TG小鼠。这些小鼠将使我们能够确定PAD4在细胞功能中的作用。