The Role of HuR in mutant SOD1 dysregulation of VEGF mRNA Processing

HuR 在突变 SOD1 VEGF mRNA 加工失调中的作用

基本信息

批准号：
7693720
负责人：
PETER H KING
金额：
$ 30.73万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-30 至 2012-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7693720
关键词：
3&apos Untranslated Regions 5&apos Untranslated Regions Abbreviations Address Affect Amino Acid Sequence Amyotrophic Lateral Sclerosis Antibodies Antioxidants Appearance Area Attenuated BRF1 gene Binding Cell Survival Cellular Stress Cessation of life Clinical Code Complex Cultured Cells Cuprozinc Superoxide Dismutase Cytoplasm Cytoplasmic Granules Dactinomycin Degenerative Disorder Disease Doxycycline EMSA Electrophoretic Mobility Shift Assay Elements Embryo Enzyme-Linked Immunosorbent Assay Enzymes Familial Amyotrophic Lateral Sclerosis Funding Gene Expression Gene Expression Regulation Genes Genetic Grant Growth Factor Half-Life IL8 gene Immunoprecipitation In Vitro Indium Interleukin-8 Interleukins Investigation Lead Ligands Link Location Maintenance Messenger RNA Methylation Modification Molecular Motor Neuron Disease Motor Neurons Mus Muscle Mutant Strains Mice Mutation Nature Neuroblastoma Neurodegenerative Disorders PTGS2 gene Paralysed Phenotype Phosphorylation Play Polyribosomes Post-Translational Protein Processing Process Progressive Disease Property Proteins Publishing RNA RNA Binding RNA Degradation RNA Processing RNA Recognition Motif RNA Splicing RNA-Binding Proteins RNA-Protein Interaction Regulation Ribonucleoproteins Role Spinal Cord Stress Superoxide Dismutase Symptoms TIS11 protein Tetanus Helper Peptide Tetracyclines Time Toxic effect Transcript Transgenic Mice Transgenic Organisms Translations Tumor Necrosis Factor-alpha Ubiquitination Untranslated Regions Vascular Endothelial Growth Factors Visual Work alpha-tocopherol transfer protein base butyrate response factor 1 copper zinc superoxide dismutase crosslink cyclooxygenase 2 effective therapy end stage disease gain of function genetic regulatory protein in vivo in vivo Model mRNA Expression mRNA Stability motor neuron degeneration mutant novel nucleocytoplasmic transport preference protein expression protein protein interaction response superoxide dismutase 1 tissue/cell culture ultraviolet

项目摘要

DESCRIPTION (provided by applicant): Amyotrophic lateral sclerosis (ALS) is a catastrophic degenerative disease of motor neurons that inexorably leads to progressive weakness and death. Genetic depletion of vascular endothelial growth factor (VEGF) leads to selective degeneration of motor neurons, providing a direct link between VEGF and ALS. Furthermore, VEGF attenuates the phenotype of ALS mice expressing the copper-zinc superoxide dismutase (SOD)-1 mutation. Posttranscriptional regulation plays a critical role in VEGF expression, particularly under conditions of cellular stress, allowing an adaptive response to promote cell survival. We recently showed that mutant SOD1 disrupts VEGF posttranscriptional regulation, leading to a significant decline in RNA and protein expression. This finding supports the toxic gain of function hypothesis for SOD1-associated ALS. These initial observations, funded by an exploratory R21 grant and recently published (Lu et al., 2007), have led to three important findings that begin to delineate the mechanism for this toxic effect. First, we discovered that mutant SOD1 is capable of directly binding AU-rich elements (ARE) in the 3' untranslated region (3'UTR) of VEGF. These cis elements are critical for posttranscriptional regulation of VEGF through interaction with cellular factors. Second, we have found that mutant but not wild-type SOD1 associates with HuR, a major regulator of VEGF mRNA stability via the ARE. Third, mutant SOD1 leads to cytoplasmic translocation and posttranslational modification of HuR. Based on these findings, we hypothesize in this proposal that mutant SOD1 exerts its negative effect on VEGF mRNA processing by disrupting the normal regulatory function of HuR. We propose three specific aims: 1) To investigate the novel RNA binding property of mutant SOD1 and its negative effect on HuR. 2) To investigate whether mutant SOD1, through its protein-protein or protein-RNA interaction, disrupts the RNA stabilizing function of HuR, or increases the association of VEGF mRNA with translational silencers or RNA destabilizers. 3) To investigate how mutant SOD1 induces HuR translocation to the cytoplasm and its post-translational modification. The long term objective of this proposal is: a) to determine how mutant SOD1 disrupts VEGF posttranscriptional regulation and b) the impact of this novel function on the ALS phenotype. PUBLIC HEALTH RELEVANCE: Amyotrophic lateral sclerosis is a relentless disease of motor neurons that leads to progressive paralysis of the muscles and ultimately death. There is no cure for this disease or any mitigating therapy. This proposal will address a novel area of growth factor regulation that may contribute to the cause of this disease, and thus may ultimately lead to novel therapies.

描述（由申请人提供）：肌萎缩性侧索硬化症（ALS）是运动神经元的灾难性退化性疾病，它无可避免地导致渐进的无力和死亡。血管内皮生长因子（VEGF）的遗传耗竭导致运动神经元的选择性变性，从而在VEGF和ALS之间提供了直接联系。此外，VEGF减轻了表达铜锌超氧化物歧化酶（SOD）-1突变的ALS小鼠的表型。转录后调节在VEGF表达中起着至关重要的作用，尤其是在细胞应激条件下，从而使自适应反应促进细胞存活。我们最近表明，突变体SOD1破坏了VEGF的转录后调节，导致RNA和蛋白质表达显着下降。该发现支持SOD1相关ALS功能假设的毒性增益。这些最初的观察结果由探索性R21赠款和最近发表（Lu等，2007）资助，导致了三个重要发现，开始描述这种有毒作用的机制。首先，我们发现突变体SOD1能够在VEGF的3'未翻译区域（3'UTR）中直接结合富含Au的元素（IS）。这些顺式元素对于通过与细胞因子的相互作用而对VEGF的转录后调节至关重要。其次，我们发现突变体而不是野生型SOD1与HUR相关，Hur是VEGF mRNA稳定性的主要调节剂。第三，突变体SOD1导致HUR的细胞质易位和翻译后修饰。基于这些发现，我们在这一建议中假设突变体SOD1通过破坏HUR的正常调节功能来对VEGF mRNA处理产生负面影响。我们提出了三个具体目标：1）研究突变体SOD1的新型RNA结合特性及其对HUR的负面影响。 2）要研究突变体SOD1是否通过其蛋白质 - 蛋白质或蛋白RNA相互作用来破坏HUR的RNA稳定功能，或增加VEGF mRNA与转化消音器或RNA DeStabilizers的关联。 3）研究突变体SOD1如何诱导HUR转移到细胞质及其翻译后修饰。该提案的长期目标是：a）确定突变体SOD1如何破坏VEGF的转录后调节，b）这种新功能对ALS表型的影响。公共卫生相关性：肌萎缩性侧索硬化症是一种无情的运动神经元疾病，导致肌肉逐渐瘫痪，并最终导致死亡。无法治愈这种疾病或任何缓解疗法。该建议将解决一个新的生长因子调节领域，该领域可能导致这种疾病的原因，因此最终可能导致新的疗法。