Polyadenylation and Translational Control

多聚腺苷酸化和翻译控制

基本信息

批准号：
10092164
负责人：
Joel D Richter
金额：
$ 47.91万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
1992
资助国家：
美国
起止时间：
1992-02-01 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10092164
关键词：
3&apos Untranslated Regions Ablation Animal Behavior Behavior Binding Brain Cell Nucleus Chemicals Complex Cytoplasm Data Defect Dendrites Disease Elements Enzymes Equilibrium Exhibits Genetic Translation Goals Hippocampus (Brain)Immunoprecipitation Impairment Injections Knockout Mice Learning Length Long-Term Depression Long-Term Potentiation Maintenance Measurement Mediating Memory Messenger RNA Metabolism Methods Modeling Molecular Multiprotein Complexes Mus Neurons Neurophysiology - biologic function Phosphorylation Physiological Poly A Poly(A) Tail Polyadenylation Polynucleotide Adenylyltransferase Pre-mRNA Polyadenylation Factor Protein Biosynthesis Proteins RNA RNA-Binding Proteins Regulation Signal Transduction Synapses Synaptic plasticity System Tail Testing Translations Work behavioral deficiency cognitive function crosslink mRNA Expression neuroregulation proteostasis ribosome profiling small hairpin RNA synaptic function transcriptome sequencing vector

项目摘要

Cytoplasmic polyadenylation is a prominent and evolutionarily conserved mechanism to regulate mRNA translation. Poly(A) length control is mediated by several factors including the RNA binding protein CPEB1, the non-canonical poly(A) polymerase Gld2, the deadenylating enzyme PARN, and the multi-protein complex CPSF. Current models indicate that CPEB1 binds the 3'UTR cytoplasmic polyadenylation element (CPE), which anchors the other proteins to RNA. PARN activity is robust and shortens the poly(A) tail, leading to translational inactivation. Upon signal-induced CPEB1 phosphorylation, PARN is expelled from the complex allowing Gld2 to catalyze polyadenylation by default, which in turn induces translation. These and other polyadenylation factors reside in neuronal dendrites where, upon synaptic stimulation, they promote polyadenylation-induced translation and resulting synaptic plasticity, the underlying cellular basis for learning and memory. Indeed. CPEB1 ablation in the brain results in synaptic impairment and behavioral deficiencies. Surprisingly, Gld2 ablation has no observable effect on animal behavior. However, a second non-canonical poly(A) polymerase, Gld4, which like Gld2 is tethered to RNA by CPEB1 but regulates the polyadenylation of different sets of mRNAs, is also present in the brain. Stereotactic injection of AAV9 vectors expressing shRNAs for Gld2 or Gld4 into the hippocampus of mice elicits little change in animal behavior. However, a double depletion of both Gld2 and Gld4 results in robust changes in behavior. These data suggest that the combination of Gld2 and Gld4 are necessary for cognitive function. We will dissect the mechanisms by which these two RNA modifying enzymes regulate mRNA metabolism and how they control neural function. This work will take the bottom-up approach of dissecting molecular mechanism, but will also help define how the brain utilizes polyadenylation and translational control to maintain proper synaptic efficacy. This work has important implications for brain activity, particularly learning and memory, and diseases associated with impairment of higher cognitive function.

细胞质聚腺苷酸化是一种突出的，进化保守的机制，用于调节mRNA 翻译。 poly（a）长度控制是由几个因素介导的，包括RNA结合蛋白CPEB1，非典型的聚（A）聚合酶GLD2，去甲基酶PARN和多蛋白质复合物 CPSF。当前模型表明CPEB1结合了3'UTR细胞质聚腺苷酸化元件（CPE），将其他蛋白锚定在RNA上。 parn活性很健壮，缩短了聚（A）的尾巴，导致翻译失活。信号诱导的CPEB1磷酸化后，将PARN从复合物中排出允许GLD2默认催化聚腺苷酸化，从而诱导翻译。这些和其他多腺苷酸化因子位于神经元树突中，在突触刺激下，它们会促进多腺苷酸化引起的翻译和产生的突触可塑性，这是学习的基础基础和内存。的确。大脑中的CPEB1消融会导致突触障碍和行为缺陷。令人惊讶的是，GLD2消融对动物行为没有明显的影响。但是，第二个非典型的聚（A）聚合酶GLD4，像GLD2一样，由CPEB1束缚在RNA上，但调节大脑中也存在不同的mRNA集合。表达AAV9载体的立体定向注入 GLD2或GLD4的shRNA进入小鼠海马，引起动物行为的变化几乎没有变化。但是， GLD2和GLD4的双重耗竭会导致行为的强大变化。这些数据表明 GLD2和GLD4的组合对于认知功能是必需的。我们将阐述这两个RNA修饰酶调节mRNA代谢以及它们如何控制神经功能。这工作将采取自下而上的方法来剖析分子机制，但也将有助于定义如何大脑利用聚腺苷酸化和转化控制来维持适当的突触功效。这项工作有对大脑活动，尤其是学习和记忆的重要含义，以及与之相关的疾病较高认知功能的损害。