Characterizing the mechanism of mitochondrial fission

线粒体裂变机制的表征

基本信息

批准号：
7156519
负责人：
Laura L Lackner
金额：
$ 4.6万
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/7156519
关键词：
Saccharomyces cerevisiae cryoelectron microscopy dynamin enzyme complex genetic regulation guanosinetriphosphatases high throughput technology inhibitor /antagonist intermolecular interaction membrane activity membrane fusion membrane reconstitution /synthesis mitochondria mitochondrial membrane molecular assembly /self assembly morphology nucleotides peptide library postdoctoral investigator protein protein interaction protein purification protein structure function small molecule

Saccharomyces cerevisiae cryoelectron microscopy dynamin enzyme complex genetic regulation guanosinetriphosphatases high throughput technology inhibitor /antagonist intermolecular interaction membrane activity membrane fusion membrane reconstitution /synthesis mitochondria mitochondrial membrane molecular assembly /self assembly morphology nucleotides peptide library postdoctoral investigator protein protein interaction protein purification protein structure function small molecule

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项目摘要

DESCRIPTION (provided by applicant): 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 Fis1 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 hvdrolysis bv Dnm1 in fission.

描述（由申请人提供）：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，因为裂变和融合蛋白已与调节凋亡的调节和人类融合蛋白的突变有关，与神经退行性疾病有关。在酿酒酵母中，至少三种蛋白对于线粒体裂变至关重要：DNM1，MDV1和FIS1有趣的是，这些蛋白质中的两种蛋白质的直系同源物中存在于哺乳动物细胞中，表明下属的分子过程是下层线粒体液体静脉静脉静脉静脉液体的分子过程。最近在酵母的研究表明，与动力蛋白相关的GTPase DNM1通过自组装和构象变化在裂变过程中驱动膜收缩，并且DNM1结构中的这些变化可能受MDV1和FIS1的调节。要详细了解DNM1，MDV1和FIS1如何组装到裂变胜任的机器中，一旦构造机器，蛋白质如何共同起作用，以介导线粒体的收缩和最终分裂，线粒体裂变事件将在体外重新构成体外。这将包括对MDV1和FIS1存在的纯化蛋白质，DNM1组装和GTPase分析的流体动力分析，以及在脂质双层上重构裂变机。这些研究将定义最小的裂变机，并将确定每种裂变蛋白以及核苷酸结合和HVDROLISSISS BV DNM1在裂变中的作用。