AMPylation, a novel mechanism regulating visual neurotransmission

AMPylation，一种调节视觉神经传递的新机制

基本信息

批准号：
8716764
负责人：
Helmut J Kramer
金额：
$ 31.16万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-01 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8716764
关键词：
Acute Adenosine Binding Proteins Biochemical Biochemical Genetics Biochemistry Biological Assay Brain Cells Data Defect Development Disease Drosophila genus Electron Microscopy Environment Enzymes Eukaryota Eukaryotic Cell Future Gene Proteins Gene-Modified Genes Genetic Genome Goals Guanosine Triphosphate Phosphohydrolases Health Immunofluorescence Microscopy Modeling Molecular Molecular Target Monomeric GTP-Binding Proteins Mutation Nervous system structure Neuroglia Pattern Phenotype Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Photoreceptors Physiological Physiological Processes Play Post-Translational Protein Processing Process Proteins Regulation Role Signal Pathway Site Staging Sterols Supraoptic Vertical Ophthalmoplegia Synapses Tertiary Protein Structure Testing Visual Visual system structure Work cell type citrate carrier fly gain of function loss of function mutant neurotransmission novel research study response rho sterol homeostasis visual process visual processing

Acute Adenosine Binding Proteins Biochemical Biochemical Genetics Biochemistry Biological Assay Brain Cells Data Defect Development Disease Drosophila genus Electron Microscopy Environment Enzymes Eukaryota Eukaryotic Cell Future Gene Proteins Gene-Modified Genes Genetic Genome Goals Guanosine Triphosphate Phosphohydrolases Health Immunofluorescence Microscopy Modeling Molecular Molecular Target Monomeric GTP-Binding Proteins Mutation Nervous system structure Neuroglia Pattern Phenotype Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Photoreceptors Physiological Physiological Processes Play Post-Translational Protein Processing Process Proteins Regulation Role Signal Pathway Site Staging Sterols Supraoptic Vertical Ophthalmoplegia Synapses Tertiary Protein Structure Testing Visual Visual system structure Work cell type citrate carrier fly gain of function loss of function mutant neurotransmission novel research study response rho sterol homeostasis visual process visual processing

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项目摘要

Cells must be able to quickly respond to changes in their environment. An important aspect of many such responses is the fast and reversible modification of proteins to regulate their function. The transient phosphorylation of proteins through the combined action of kinases and phosphatases may be the most prominent example of such regulation. A similar, more recently discovered modification of proteins is the stabile addition of adenosine 5'--‐monophosphate group (AMP) to proteins. This novel regulatory mechanism has been called AMPylation after its initial discovery in the context of the bacterial VopS protein. Since then, bacterial and eukaryotic Fic domains have been found that can AMPylate proteins. Drosophila offers a significant advantage for the analysis of the physiological role of AMPylation because fly genomes encode only a single Fic domain protein. The specific aims described in this proposal combine biochemical and genetic approaches in Drosophila to define the role that this mechanism plays in visual neurotransmission. This proposal aims (i) to determine the cellular sites of AMPylation activity using immunofluorescence and electron microscopy approaches, (ii) to define developmental stages and cell types that require AMPylation, (iii) to analyze the physiological consequences of AMPylation in the context of the Drosophila visual system, and (iv) to employ biochemical purification in conjunction with genetic interaction assays to identify the molecular targets regulated by AMPylation. Completion of these experiments will yield a comprehensive understanding of the signaling pathways in the visual system that are modified by AMPylation.

细胞必须能够快速响应其环境变化。许多此类反应的一个重要方面是对蛋白质调节其功能的快速和可逆修饰。蛋白质通过激酶和磷酸酶的联合作用的瞬时磷酸化可能是这种调节的最突出的例子。最近发现的类似的蛋白质修饰是稳定腺苷5' - 单磷酸基（AMP）与蛋白质的稳定性。在细菌VOPS蛋白的背景下，这种新型的调节机制在其初始发现后被称为重酰化。从那时起，就发现细菌和真核FIC结构域可以对聚青铜蛋白。果蝇为分析两极的生理作用提供了重要的优势，因为蝇基因组仅编码单个FIC结构域蛋白。该提案中描述的具体目的结合了果蝇中的生化和遗传方法，以定义该机制在视觉神经传递中的作用。该提议旨在（i）使用免疫荧光和电子显微镜方法来确定两极分化活性的细胞位点（ii），以定义需要两极化的发育阶段和细胞类型，（iii）分析在drosophiala visial contion concition concition conction conction conction con CONSITION中的生理后果，并（IV）的生理后果分子靶标由两臂调节。这些实验的完成将对视觉系统中的信号传导途径进行全面的了解，这些信号通路已通过两支化合物修饰。