Neuronal function of huntingtin associated protein

亨廷顿相关蛋白的神经元功能

基本信息

批准号：
7213013
负责人：
XIAO-JIANG LI
金额：
$ 33.47万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-01-01 至 2011-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7213013
关键词：
14-3-3 Proteins ART protein ATP phosphohydrolase Adaptor Signaling Protein Address Adenovirus Vector Adenoviruses Adult Affect Afferent Neurons Age Alanine Alleles Alternative Splicing Amino Acids Amygdaloid structure Animal Behavior Animal Model Animals Antibodies Antigens Aprotinin Area Axon Axonal Transport Bacteria Behavioral Bilateral Binding Binding Sites Biochemical Biological Biological Assay Biotin Biotinylation Birth Body Size Body Weight Body Weight decreased Brain Brain region Brain-Derived Neurotrophic Factor Breeding Buffers Bypass C-terminal Calcium/calmodulin-dependent protein kinase California Catalytic Domain Cause of Death Cavia Cell Count Cell Fraction Cell Fractionation Cell Nucleus Cell Survival Cell membrane Cell model Cell physiology Cell surface Cells Centrifugation Cessation of life Chimeric Proteins Cleaved cell Cloning Code Collaborations Complementary DNA Complex Computer software Condition Confocal Microscopy Contralateral Corpus striatum structure Culture Media Cultured Cells Cyclic AMP Cyclic AMP-Dependent Protein Kinases Cytolysis Cytomegalovirus Cytoplasm Cytoplasmic Inclusion Cytosol Data Defect Densitometry Dependovirus Depth Diet Diffuse Digestion Disease Disruption Distal Drosophila genus Dyes Dynein ATPase ES Cell Line Eating Eating Behavior Egtazic Acid Electron Microscopy Employee Strikes Endocytosis Endosomes Ensure Epidermal Growth Factor Epidermal Growth Factor Receptor Epitopes Equilibrium Event Exons Family Fc Receptor Feeding behaviors Female Fetus Figs - dietary Fluorescence Fluorescence Recovery After Photobleaching Fluorescent Dyes Food Forelimb Fractionation Freezing Functional disorder Funding Future GABA Receptor Gamma counter Gene Targeting Generations Genes Genetic Genetic Recombination Genomics Genotype Gills Glass Glues Glutamates Glutamine Glutathione Glutathione S-Transferase Glycerophosphates Goals Golgi Apparatus Grant Green Fluorescent Proteins Growth Growth Factor Receptors Hand Harvest Head Hindlimb Hippocampus (Brain)Homologous Gene Human Human Amyloid Precursor Protein Huntington Disease Hypothalamic structure Ice Image Image Analysis Immunoblotting Immunoglobulin G Immunoprecipitation Impairment In Vitro Incubated Individual Infection Injection of therapeutic agent Inositol Institutes Intake Intracellular Transport Investigation Japan Ketamine Kinesin Kinetics Knock-in Mouse Knock-out Knockout Mice Label Laboratories Lead Left Length Letters Leupeptins Life Ligand Binding Ligands Ligation Ligature Light Link Localized Long-Term Effects Lysine Lysosomes Mammalian Cell Manuscripts Masks Measurement Measures Mediating Mediator of activation protein Medicine Membrane Membrane Protein Traffic Membrane Proteins Memory Loss Methods Microinjections Microscope Microscopy Microtubules Mitochondria Modeling Molecular Monitor Monoclonal Antibodies Morphology Motor Motor Neuron Disease Motor Neurons Movement Movement Disorders Multivesicular Body Mus Mutate Mutation N-terminal NGFR Protein Nature Neomycin Neomycin resistance gene Nerve Nerve Degeneration Nerve Growth Factors Neuraxis Neurites Neuritis Neurodegenerative Disorders Neuroglia Neurologic Neurologist Neuronal Differentiation Neuronal Dysfunction Neurons Neurosciences Neurotrophic Tyrosine Kinase Receptor Type 1 Newborn Animals Nonidet P-40 Nuclear Numbers Nycomed Okadaic Acid Organelles Oryctolagus cuniculus Outcome PC12 Cells PKA inhibitor POMC gene Paper Partner in relationship Pathogenesis Pathology Pathway interactions Patients Peptides Peripheral Personal Satisfaction Pharmaceutical Preparations Phenotype Philadelphia Phosphate Buffer Phosphoric Monoester Hydrolases Phosphorylation Phosphorylation Site Phosphotransferases Photobleaching Plasmids Play Polymerase Chain Reaction Post-Translational Protein Processing Precipitation Preparation Principal Investigator Pro-Opiomelanocortin Procedures Process Progress Reports Promega Protein Binding Protein Conformation Protein Isoforms Protein Kinase Protein Phosphorylation Protein Overexpression Proteins Proto-Oncogene Proteins c-akt Protocols documentation Publications Publishing Purpose Rate Rattus Reaction Reagent Receptor Signaling Recombinant Proteins Recovery Recycling Regulation Relative (related person)Reporter Reporting Research Research Personnel Resolution Right-On Rodent Role Rotarod Performance Test Sampling Scaffolding Protein Screening procedure Serine Serum-Free Culture Media Signal Pathway Signal Transduction Signaling Molecule Site Site-Directed Mutagenesis Slide Small Interfering RNA Solid Solutions Southern Blotting Specificity Spinal Ganglia Staging Staining method Stains Standards of Weights and Measures Staurosporine Streptavidin Stretching Stroke Structure Structure of nucleus infundibularis hypothalami Subcellular Fractions Subcellular structure Sucrose Surface Symptoms Synaptic Vesicles System Tail Testing Texas red Thalamic structure Thigh structure Thinking Threonine Thymidine Kinase Time Tissues Toxic effect Tracer Transfection Transferrin Transgenes Transgenic Mice Transgenic Organisms Transport Vesicles TrkA protein Tubulin United States National Institutes of Health Vertebrates Vesicle Viral Viral Vector Virus Virus Diseases Water Water consumption Week Western Blotting Wild Type Mouse Work Yang Yeasts analog anterograde transport antibody conjugate base biotin 1 blastocyst brain tissue calmodulin-dependent protein kinase II cell growth regulation cell type cellular pathology cryostat cytotoxicity day design dimer dosage dynactin dynactin 1 embryonic stem cell enolase experience factor A feeding fluorescence imaging gain of function gang homologous recombination human APEX1 protein human Huntingtin protein human disease immunocytochemistry immunoreaction in vivo insight instrument interest intracellular protein transport knock-down leupeptin loss of function male microbial alkaline proteinase inhibitor mouse Hap1 protein mouse model movie mutant neuronal cell body neuronal survival neuronal transport neuropathology neuropeptide Y2 receptor neurotoxicity numb protein particle phosphatase inhibitor plasmid DNA polyclonal antibody polyfunctional aziridine polyglutamine polypeptide postnatal programs promoter protein aggregation protein transport pup rat Hap1 protein receptor receptor expression receptor function receptor internalization receptor recycling recombinase relating to nervous system research study response retrograde transport sample fixation sciatic nerve sulfo-N-hydroxysuccinimide-biotin synaptic function theories trafficking transcriptional coactivator p75 transmission process trend tripolyphosphate tyrosine kinase ABL1 vector xylamine yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Huntingtin associated protein-1 (HAP1) is enriched in neurons and is found to be involved in intracellular trafficking. The role of HAP1 in intracellular trafficking is supported by its interactions with a number of proteins including dynactin p150 and kinesin, which are involved in microtubule-dependent transport. Recent studies also suggest that HAP1 participates in vesicular trafficking and endocytosis of membrane receptors. The crucial function of HAP1 was further demonstrated by HAP1 knockout mice that are postnatally lethal and show neurodegeneration in the brain. These interesting findings raise more questions about how HAP1 is involved in the complex intracellular trafficking in neurons. Our recent studies show that HAP1 interacts with 14-3-3 and that this interaction is regulated by phosphorylation of HAP1. We hypothesize that HAP1 may link specific cargos to the microtubule transporters and that it's interactions with partners are regulated by its posttranslational modifications. Such regulation allows HAP1 to participate in intracellular transport of various cargos and endocytosis of membrane receptors. Given the idea that HAP1 dysfunction may be involved in HD pathology, it is also important to investigate if the loss of HAP1 can affect its function in intracellular trafficking. Accordingly, we propose three aims in this application. In Aim 1, we will investigate whether 14-3-3 and phosphorylation regulate the interactions of HAP1 with microtubule transporters dynactin p150 and kinesin. In Aim-2, we will use HAP1 knockout mice and siRNA approaches to examine whether decreasing MAPI's expression alters microtubule-dependent transport and neuronal function in different types of neurons. In Aim-3, we will study whether HAP1 deficiency affects endocytosis of membrane receptors and whether defective HAP1-associated endocytosis contributes to HD pathology. These studies aim to elucidate the function of HAP1 and to help understand the mechanisms for intracellular trafficking in neurons.

描述（由申请人提供）：亨廷顿蛋白相关蛋白 1 (HAP1) 在神经元中富集，并被发现参与细胞内运输。 HAP1 在细胞内运输中的作用得到了其与许多蛋白质的相互作用的支持，包括动力蛋白 p150 和驱动蛋白，这些蛋白质参与微管依赖性运输。最近的研究还表明，HAP1 参与膜受体的囊泡运输和内吞作用。 HAP1 基因敲除小鼠进一步证明了 HAP1 的关键功能，这些小鼠在出生后是致命的，并表现出大脑神经退行性变。这些有趣的发现提出了更多关于 HAP1 如何参与神经元复杂的细胞内运输的问题。我们最近的研究表明，HAP1 与 14-3-3 相互作用，并且这种相互作用受到 HAP1 磷酸化的调节。我们假设 HAP1 可能将特定的货物与微管转运蛋白连接起来，并且它与伙伴的相互作用受到其翻译后修饰的调节。这种调节使得HAP1能够参与细胞内各种货物的运输和膜受体的内吞作用。鉴于 HAP1 功能障碍可能与 HD 病理有关，因此研究 HAP1 的丢失是否会影响其在细胞内运输中的功能也很重要。因此，我们在此应用中提出了三个目标。在目标 1 中，我们将研究 14-3-3 和磷酸化是否调节 HAP1 与微管转运蛋白 dynactin p150 和驱动蛋白的相互作用。在 Aim-2 中，我们将使用 HAP1 敲除小鼠和 siRNA 方法来检查降低 MAPI 的表达是否会改变不同类型神经元中的微管依赖性运输和神经元功能。在 Aim-3 中，我们将研究 HAP1 缺陷是否影响膜受体的内吞作用，以及 HAP1 相关内吞作用缺陷是否会导致 HD 病理。这些研究旨在阐明 HAP1 的功能并帮助了解神经元细胞内运输的机制。