Upstream open reading frames in neuronal function: a singular and genome-wide approach

神经元功能中的上游开放阅读框：单一的全基因组方法

• 批准号: 8837820
• 负责人: Caitlin Marie Rodriguez
• 金额: $ 3.34万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8837820
• 关键词: 5' Untranslated Regions; 6-Cyano-7-nitroquinoxaline-2,3-dione; Agonist; Area; Autistic Disorder; Biochemical; Bioinformatics; Biological Models; Biology; CGG repeat; Case Study; Cell Line; Clinical; Codon Nucleotides; Data; Disease; Emerging Technologies; Event; FMR1; FMR1 Gene; FMR1 repeat; FMRP; FXTAS; Fragile X Syndrome; Future; Gene Expression Profile; Genes; Glycine; Goals; Grant; Intellectual functioning disability; Intervention; Lead; Learning; Maps; Measures; Mediating; Memory; Messenger RNA; Nerve Degeneration; Neurobiology; Neurodegenerative Disorders; Neurodevelopmental Disorder; Neurons; Nucleotides; Open Reading Frames; Pathogenesis; Play; Positioning Attribute; Prevalence; Process; Production; Protein Biosynthesis; Proteins; RNA; Reporter; Research; Ribosomes; Role; Scanning; Series; Site; Stream; Structure; Synaptic plasticity; Syndrome; Techniques; Testing; Training; Transcript; Translating; Translational Regulation; Translations; Untranslated Regions; Work; base; career; cellular imaging; channel blockers; genome-wide; insight; kainate; nervous system disorder; neuroblastoma cell; next generation sequencing; novel; public health relevance; response; synaptic function

 DESCRIPTION (provided by applicant): Neurons regulate when and where they synthesize new proteins to maintain basal and activity-dependent compartmental fidelity. Control of this process is largely regulated at translational initiation which typically occurs at the 5'-most AUG codon in an appropriate mRNA sequence context. Emerging data in simple model systems suggests that un-annotated, upstream open reading frames (uORFs) are utilized on a significant fraction of all transcripts. These uORFs often initiate at near-AUG (one base different from AUG) codons and display condition-dependent usage, suggesting they may play a regulatory role in translational control. How such atypical initiation events are mediated and what roles they play in neuronal function and disease are unknown. The central hypothesis of this grant is that upstream open reading frame usage is an important regulator of translational dynamics in neurons, and that proteins produced by translation from un-annotated uORFs will contribute to neuronal function. Our group recently discovered a novel mechanism for protein translational initiation associated with nucleotide repeats in the neurodegenerative disorder Fragile X-associated Tremor Ataxia Syndrome (FXTAS). FXTAS results from a CGG nucleotide repeat expansion in the 5' untranslated region (UTR) of the FMR1 gene. We discovered that this repeat expansion elicits initiation at a near-AUG codon in the 5'UTR, resulting in translation through the repeat via a process known as RAN (Repeat Associated Non-AUG) translation. This translation event leads to synthesis of a poly-glycine containing protein, FMRpolyG, even at normal repeat sizes. This proposal will first determine when FMRpolyG is synthesized in neurons both basally and in response to activity. Further, the impact of FMRpolyG synthesis on translation of the major FMR1 gene product, FMRP- a protein implicated in autism and intellectual disability will be assessed. To determine the prevalence of similar uORF utilization events in neurons, the novel next-generation sequencing technique known as ribosomal profiling will be used to measure ribosomal occupancy within 5'UTRs across the neuronal transcriptome. These studies will employ cellular imaging, biochemical and bioinformatics techniques to explore a new area in neurobiology with relevance to both synaptic plasticity as well as numerous neurological diseases.

 描述（由申请人证明）：何时和乳清合成新的蛋白质，以维持所有转录本的数量。 - 依赖的用法，他们在翻译控制中起着监管作用。 在神经元功能和疾病中的发挥是，上游的开放式阅读是神经元转化动力学的重要调节器，并且通过未经通知的UORF的翻译产生的蛋白质将有助于我们小组的神经元功能。 NeuroDeatirderater脆弱的X相关性共济失调综合征（FXTA）。 通过称为RAN的过程（重复ASCIAND NON-AUG）翻译。 FMRPOLYG合成对自闭症和智力障碍的主要FMR1基因产物的翻译的影响将被评估。 5'Utrss l转录组。