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：
8309929
负责人：
Helmut J Kramer
金额：
$ 31.8万
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/8309929
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

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环境的变化。许多此类反应的一个重要方面是蛋白质的快速且可逆的修饰以调节其功能。通过激酶和磷酸酶的联合作用实现的蛋白质瞬时磷酸化可能是这种调节的最突出的例子。最近发现的一种类似的蛋白质修饰是在蛋白质上稳定添加 5'--单磷酸腺苷 (AMP)。这种新颖的调节机制在细菌 VopS 蛋白中首次发现后被称为 AMPylation。从那时起，细菌和真核生物的 Fic 结构域被发现可以 AMPylate 蛋白质。果蝇为分析 AMPylation 的生理作用提供了显着的优势，因为果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标结合了果蝇中的生化和遗传学方法，以确定该机制在视觉神经传递中发挥的作用。该提案的目的是（i）使用免疫荧光和电子显微镜方法确定 AMPylation 活性的细胞位点，（ii）定义需要 AMPylation 的发育阶段和细胞类型，（iii）分析 AMPylation 在果蝇视觉系统，以及 (iv) 采用生化纯化结合遗传相互作用测定来识别 AMPylation 调节的分子靶标。完成这些实验将全面了解视觉系统中受 AMPylation 修饰的信号通路。