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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和可逆修饰以调节其功能。通过激酶和磷酸酶的联合作用，蛋白质的瞬时磷酸化可能是这种调节的最突出的例子。最近发现的一种类似的蛋白质修饰是向蛋白质中稳定地添加腺苷5 '-一磷酸基团（AMP）。这种新的调节机制在细菌VopS蛋白的背景下首次发现后被称为AMP化。从那时起，已经发现细菌和真核生物的Fic结构域可以使蛋白质腺苷酸化。果蝇为分析AMP化的生理作用提供了显著的优势，因为果蝇基因组仅编码单个Fic结构域蛋白。在这个建议中描述的具体目标联合收割机结合果蝇的生物化学和遗传学的方法来定义的作用，这种机制在视觉神经传递。该建议旨在（i）使用免疫荧光和电子显微镜方法确定AMP化活性的细胞位点，（ii）确定需要AMP化的发育阶段和细胞类型，（iii）分析果蝇视觉系统中AMP化的生理后果，和（iv）采用生物化学纯化结合遗传相互作用测定来鉴定由AMP化调节的分子靶标。这些实验的完成将产生一个全面的理解，在视觉系统中的信号通路被修改的AMP。