3-D FINE STRUCTURE OF MULTIVESICULAR BODIES IN SACCHAROMYCES CEREVISIAE

酿酒酵母多胞体的 3-D 精细结构

• 批准号: 8170819
• 负责人: CHARLES G ODORIZZI
• 金额: $ 2.49万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170819
• 关键词: 3-Dimensional; Back; Biochemical; Cell Proliferation; Cell Surface Receptors; Cell membrane; Cells; Computer Retrieval of Information on Scientific Projects Database; Cytoplasm; Cytoplasmic Protein; Destinations; Endocytosis; Endosomes; Eukaryotic Cell; Funding; Future; Genes; Genetic; Genetic Screening; Grant; Homologous Gene; Human; Hydrolase; Institution; Lysosomes; Membrane; Morphology; Multivesicular Body; Mutation; Pathway interactions; Proteins; Recruitment Activity; Recycling; Research; Research Personnel; Resources; Role; Route; Saccharomyces cerevisiae; Saccharomycetales; Shapes; Sorting - Cell Movement; Source; Structure; Testing; Time; Tomogram; United States National Institutes of Health; Vesicle; Work; human disease; macromolecule; novel; therapeutic target; yeast protein

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The endocytic pathway functions to segregate ingested macromolecules that are destined to be degraded in the lysosome from molecules that are either recycled back to the plasma membrane or routed toward other intracellular destinations. One of the most recognizable endocytic compartments is the multivesicular body (MVB), which contains within its lumen membrane-enclosed vesicles that are formed by inward budding of the limiting endosomal membrane. Many activated cell-surface receptors that control cell proliferation are down-regulated through endocytosis and sorting into the lumenal vesicles of MVBs. These vesicles and their contents are subsequently degraded by lysosomal hydrolases upon fusion of the limiting MVB membrane with the lysosome. Preliminary tomograms have shown that wild-type budding yeasts contain MVBs that are similar to those in higher eukaryotic cells. A novel genetic screen was used to uncover the BRO1 gene, which encodes a conserved, soluble cytoplasmic protein that associates with endosomal membranes. The Bro1 protein may be recruited from the cytoplasm to the endosome in order to coordinate inward invagination of the endosomal membrane and budding of vesicles toward the compartment's lumen. To test this hypothesis, the function and localization of Bro1 proteins that have domain-specific mutations will be analyzed. Initially, we completed 9 dual-axis tomograms of BRO1-deleted cells and found that these cells contained aberrant membrane compartments with three distinct morphologies. These included stacked cisternae, stacked cisternae that were curved into a C-shape, and curved cisternae enclosing a spherical compartment. Future studies will use genetic and biochemical analysis to identify proteins that cooperate with the wild-type Bro1 protein in MVB vesicle formation. The Odorizzi lab has recently hired a full time technician, Matthew West who is using the Resource facilities to work on additional strains that have mutations in other genes required for MVB sorting. Together, these studies should define the functional role of the Bro1 protein in MVB vesicle formation and identify new yeast proteins with human homologues that could serve as targets for therapeutics in human diseases.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 内吞途径的功能可将注定要在溶酶体中降解的摄入的大分子从分子中降解，这些分子可以回收回到质膜，或者朝其他细胞内目的地延伸。 最知名的内吞区室之一是多囊体（MVB），其中包含其腔膜封闭的囊泡，这些囊泡是由限制内体膜向内萌芽形成的。 许多控制细胞增殖的活化的细胞表面受体通过内吞作用并分类为MVBS的腔囊泡。 这些囊泡及其含量随后被溶酶体水解酶与裂解膜融合后降解。 初步断层图表明，野生型萌芽的酵母包含与较高真核细胞相似的MVB。一个新型的遗传筛选被用来揭示BRO1基因，该基因编码与内体膜相关的保守的可溶性细胞质蛋白。 BRO1蛋白可以从细胞质募集到内体募集，以协调内体膜的内向内，囊泡向腔室的腔内萌芽。 为了检验该假设，将分析具有域特异性突变的BRO1蛋白的功能和定位。 最初，我们完成了9个骨骼骨骼细胞的9个双轴断层图，发现这些细胞包含具有三种不同形态的异常膜室。其中包括堆叠的蓄水池，堆叠的蓄水池，弯曲成C形，并弯曲的水箱封闭了球形隔室。未来的研究将使用遗传和生化分析来鉴定与MVB囊泡形成中野生型BRO1蛋白合作的蛋白质。 Odorizzi Lab最近聘请了一名全职技术人员Matthew West，他正在使用资源设施来处理其他菌株，这些菌株在MVB分类所需的其他基因中具有突变。 总之，这些研究应定义Bro1蛋白在MVB囊泡形成中的功能作用，并鉴定具有人类同源物的新酵母菌蛋白，这些蛋白质可以用作人类疾病中疗法的靶标。