Phospholipid Dynamics in Membrane Assembly

膜组装中的磷脂动力学

基本信息

批准号：
7883194
负责人：
DENNIS R. VOELKER
金额：
$ 38.61万
依托单位：
NATIONAL JEWISH HEALTH
依托单位国家：
美国
项目类别：
财政年份：
1983
资助国家：
美国
起止时间：
1983-07-01 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7883194
关键词：
1-Phosphatidylinositol 4-Kinase ATP phosphohydrolase Acyl Coenzyme A Acylation Acyltransferase Address Affect Affinity Alanine Algorithms Alleles Anabolism Antibodies Appearance Applied Genetics Area Award Binding Binding Proteins Biochemical Biochemical Genetics Biochemistry Biogenesis Biological Biological Assay Biological Transport C2 Domain Carboxy-Lyases Carrier Proteins Catalysis Cell membrane Cells Cellular biology Centrifugation Characteristics Collaborations Commit Complement Complex Coupled Coupling Data Databases Decarboxylation Defect Deubiquitinating Enzyme Deubiquitination Dissection Docking Embryo Endoplasmic Reticulum Enzymes Ergosterol Essential Genes Ethanolamines Eukaryota Eukaryotic Cell Event Exhibits Failure Family Figs - dietary Genes Genetic Genetic Recombination Genetic Screening Genetic Transcription Glutathione Golgi Apparatus Grant Growth Heart Diseases Homeostasis Hypersensitivity In Vitro Individual Inner mitochondrial membrane Inositol Integral Membrane Protein Intoxication Investigation Isotopes Knowledge Laboratories Lecithin Lesion Lipid Binding Lipids Liposomes Location Lysophospholipids Maintenance Malignant Neoplasms Mammalian Cell Mammals Measures Membrane Membrane Proteins Membrane Transport Proteins Metabolism Methionine Mitochondria Mitochondrial Proteins Modeling Molecular Molecular Chaperones Molecular Genetics Molecular Machines Mono-S Mouse Strains Multiprotein Complexes Mus Mutation Names Nature Oligonucleotides Open Reading Frames Organelles Organism Other Genetics Pathway interactions Phase Phenocopy Phenotype Phosphatidic Acid Phosphatidyl glycerol Phosphatidylethanolamine Phosphatidylglycerols Phosphatidylinositols Phosphatidylserines Phospholipids Phosphorylcholine Plasmodium falciparum Play Process Production Protein Binding Proteins Radial Radioisotopes Reaction Regulation Relative (related person)Research Research Personnel Retrieval Role Route SKP Cullin F-Box Protein Ligases Salts Sedimentation process Series Serine Side Signal Transduction Site Solid Sorting - Cell Movement Source Stable Populations Sterol O-Acyltransferase Structure Sucrose Sulfur Sulfur Amino Acids Supplementation Surface System Testing Threonine Toxic effect Toxoplasma gondii Transmembrane Transport Transport Process Transport Reaction Ubiquitin Ubiquitin-Protein Ligase Complexes Ubiquitination Vascular Plant Western Blotting Work Yeasts acronyms aminoacid biosynthesis aminophospholipid transporter analog analytical tool base cadmium ion cell growth cell type cofactor density enzyme activity gene cloning genetic association genetic manipulation human disease in vivo insight interest lipid metabolism lipid transport membrane assembly membrane biogenesis mitochondrial dysfunction mutant phosphatidylethanolamine phosphatidylinositol 4-phosphate phosphoethanolamine methyltransferase positional cloning protein complex protein protein interaction reconstitution research study scaffold success tool toxic metal trafficking transcription factor transmission process ubiquitin ligase ubiquitin-protein ligase uptake yeast two hybrid system

项目摘要

A fundamental unsolved problem of cell biology and biochemistry is the molecular definition of the mechanisms of phospholipid transport for membrane biogenesis. This process is essential for all cell growth, replication, differentiation and homeostasis. The focus of this proposal is to use the power of yeast molecular genetics to dissect the process and understand the general principles of how it occurs throughout eukaryotic organisms. We have made progress in defining several of the genes and gene products that participate in the transport reactions, but large gaps in our knowledge still remain. The work will continue to concentrate on the transport reactions involving the aminoglycerophospholipids (phosphatidylserine, phosphatidylethanolamine and phosphatidylcholine), since the tools we have previously developed greatly facilitate measuring inter-membrane transfer events, and also provide the basis of genetic screens and selections for strains that are defective in these transport processes. In the first Specific Aim we will further define the role of protein ubiquitination in regulating physical associations between the endoplasmic reticulum and the mitochondria, and in serving as a nucleation site for the assembly of multi-protein complexes involved in moving the lipids. In the second Specific Aim we will extend our findings from previous genetic, and protein-protein interaction studies, to reconstitute the associations between biological and synthetic donor membranes in vitro; and mechanistically define how individual proteins and lipids regulate the transport of specific aminoglcyerophospholipids. In the third Specific Aim we will continue to apply genetic and biochemical tools to elucidate the genes and gene products involved in the export of phosphatidylethanolamine from mitochondria and the Golgi apparatus. In the fourth Specific Aim we will use recently developed conditional alleles for phosphatidylserine decarboxylaseexpressed in mice, to define the function of this enzyme in mitochondrial biogenesis in mammalian systems. The combined genetic and biochemical approaches we propose will provide new mechanistic and molecular information about phospholipid transport processes in eukaryotic cells and provide new insights into the regulation of membrane biogenesis. This fundamental work is most relevant to areas of human disease concerned with control of cell growth and mitochondrial dysfunction, such as cancer and heart disease.

细胞生物学和生物化学的基本未解决问题是分子的定义磷脂转运的机理用于膜生物发生。这个过程对于所有单元都至关重要生长，复制，分化和稳态。该提议的重点是使用酵母的力量分子遗传学剖析过程并了解其在整个过程中的发生的一般原理真核生物。我们在定义几种基因和基因产物方面取得了进步参与运输反应，但我们的知识仍然存在很大的差距。工作将继续集中于涉及氨基甘油磷脂（磷脂酰丝氨酸，，，，磷脂酰乙醇胺和磷脂酰胆碱），因为我们以前已经开发了大量的工具促进测量膜间转移事件，还提供了遗传筛选的基础在这些运输过程中有缺陷的菌株的选择。在第一个特定目标中，我们将进一步定义蛋白质泛素化在调节内质之间物理关联中的作用网状和线粒体，并用作组装多蛋白的成核位点涉及移动脂质的复合物。在第二个特定目标中，我们将从以前的遗传和蛋白质 - 蛋白质相互作用研究，以重建生物学之间的关联和体外合成供体膜；并机械地定义了单个蛋白质和脂质的方式调节特定氨基糖磷脂的转运。在第三个特定目标中，我们将继续应用遗传和生化工具来阐明与出口有关的基因和基因产物线粒体和高尔基体的磷脂酰乙醇胺。在第四个特定目标中，我们将使用最近开发了用于小鼠磷脂酰丝氨酸脱羧的条件等位基因，以定义该酶在哺乳动物系统中线粒体生物发生中的功能。遗传和我们提出的生化方法将提供有关有关的新机械和分子信息真核细胞中的磷脂转运过程，并为调节提供新的见解膜生物发生。这项基本工作与与人类疾病有关的领域最相关控制细胞生长和线粒体功能障碍，例如癌症和心脏病。