Characterizing the mechanism of mitochondrial fission

线粒体裂变机制的表征

基本信息

批准号：
7316866
负责人：
Laura L Lackner
金额：
$ 4.88万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-12-01 至 2009-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7316866
关键词：
Affect Apoptosis Binding Biological Assay Build-it Cell physiology Cellular biology Centrifugation Chemistry Chimeric Proteins Complex Constriction procedure Data Dynamin Eukaryota Eukaryotic Cell Event Fellowship Gel Chromatography Guanosine Triphosphate Phosphohydrolases Human Hydrolysis In Vitro Individual Institutes Link Lipid Bilayers Lipids Malignant Neoplasms Mammalian Cell Mediating Membrane Mitochondria Molecular Mutation Names Neurodegenerative Disorders Nucleotides Organelles Orthologous Gene Outer Mitochondrial Membrane Play Process Property Proteins Regulation Resolution Role Saccharomyces cerevisiae Shapes Structure Sucrose Surface System Therapeutic Work Yeasts inhibitor/antagonist insight intermolecular interaction protein function proteoliposomes reconstitution self assembly size small molecule small molecule libraries tool

Affect Apoptosis Binding Biological Assay Build-it Cell physiology Cellular biology Centrifugation Chemistry Chimeric Proteins Complex Constriction procedure Data Dynamin Eukaryota Eukaryotic Cell Event Fellowship Gel Chromatography Guanosine Triphosphate Phosphohydrolases Human Hydrolysis In Vitro Individual Institutes Link Lipid Bilayers Lipids Malignant Neoplasms Mammalian Cell Mediating Membrane Mitochondria Molecular Mutation Names Neurodegenerative Disorders Nucleotides Organelles Orthologous Gene Outer Mitochondrial Membrane Play Process Property Proteins Regulation Resolution Role Saccharomyces cerevisiae Shapes Structure Sucrose Surface System Therapeutic Work Yeasts inhibitor/antagonist insight intermolecular interaction protein function proteoliposomes reconstitution self assembly size small molecule small molecule libraries tool

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项目摘要

Mitochondria are dynamic organelles that continuously undergo fission and fusion. These dynamic events regulate the size and shape of mitochondria and play critical roles in cell physiology, as fission and fusion proteins have been implicated in the regulation of apoptosis and mutations in human fusion proteins have been linked to neurodegenerative diseases. In the yeast S. cerevisiae, at least three proteins are essential for mitochondrial fission: Dnm1, Mdv1, and Fisl Interestingly, orthologs of two of these proteins exist in mammalian cells indicating that the molecular processes underlying mitochondrial fission are well conserved. Recent studies in yeast suggest the dynamin related GTPase Dnm1 drives membrane constriction during fission by self-assembly and conformational changes, and that these alterations in Dnm1 structure may be regulated by Mdv1 and Fis1. To understand in mechanistic detail how Dnm1, Mdv1, and Fis1 assemble into a fission competent machine and, once the machine is constructed, how the proteins function together to mediate the constriction and ultimate division of mitochondria, mitochondrial fission events will be reconstituted in vitro. This will include hydrodynamic analysis of purified proteins, Dnm1 assembly and GTPase assays in the presence of Mdv1 and Fis1, and the reconstitution of the fission machine on lipid bi- layers. These studies will define the minimal fission machine and will determine the roles of each fission protein and of nucleotide binding and hvdrolvsis bv Dnm1 in fission.

线粒体是不断经历裂变和融合的动态细胞器。这些动态事件调节线粒体的大小和形状，并在细胞生理中起关键作用，作为裂变和融合蛋白质与人凋亡的调节有关，人类融合蛋白的突变已与神经退行性疾病有关。在酿酒酵母中，至少三种蛋白质是必不可少的对于线粒体裂变：DNM1，MDV1和FISL有趣的是，这些蛋白中的两种直系同源物存在于哺乳动物细胞表明线粒体裂变的分子过程是充分保守的。酵母的最新研究表明，与Dynamin相关的GTPase DNM1驱动膜收缩。通过自组装和构象变化进行裂变，DNM1结构中的这些改变可能是由MDV1和FIS1调节。了解机械的详细信息DNM1，MDV1和FIS1如何组装成裂变胜任的机器，一旦构造了机器，蛋白质如何共同发挥作用调解线粒体的收缩和最终划分，线粒体裂变事件将是在体外重构。这将包括纯化蛋白质，DNM1组装和 GTPase在存在MDV1和FIS1的情况下进行的GTPase分析，以及在脂质BI-上重构的裂变机层。这些研究将定义最小的裂变机，并确定每个裂变的作用蛋白质和核苷酸结合和HVDROLVSIS BV DNM1在裂变中。