Molecular mechanisms of endo- and exocytosis in the baker's yeast

面包酵母内吞作用和胞吐作用的分子机制

• 批准号: 249976640
• 负责人: Professor Dr. Adam Bertl
• 金额: --
• 依托单位: Arbeitsgruppe Plant Membrane Biophysics
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2019-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/249976640?language=en
• 关键词: Molecular; mechanisms; endo; exocytosis; baker

Endo- and exocytosis are imperative in numerous cellular processes in eukaryotes, such as polar growth, cell division, motility or signaling. By fusion of exocytotic vesicles cells deliver secretory cargo, membrane-resident proteins and also membrane to the plasma membrane; the retrogarde transport of these materials is provided by fisson of endocytotic vesicles. Since the electrical properties of a cell membrane are comprised of a selective conductance and a capacitance, the latter being proportional to the membrane surface area, changes in the surface area from fusion and fission of vesicles can be directly monitored by recording of the membrane capacitance. In this way the size of individual exo- and endocytotic vesicles and their kinetics of fusion and fission can be recorded in real time in living cells. After showing that this recording technique is also feasible in yeast protoplasts we will in this proposal combine the detailed genetic and biochemical knowledge on exo-/endocytosis in yeast with high resolution capacitance recordings. The use of the rich library of yeast mutants with defects in distinct proteins in the exo- and endocytotic machinery and the use of well established signals for membrane trafficking like the mating pheromones will make it possible to dissect the complex mechanisms of fusion and fission into well defined molecular steps. Because of the conservation of the molecular machinery the results of this work will have strong implications for the understanding of these processes in mammalian cells.

内吞作用和胞吐作用在真核生物的许多细胞过程中是必不可少的，例如极性生长、细胞分裂、运动或信号传导。通过胞吐囊泡的融合，细胞将分泌物、膜驻留蛋白和膜递送至质膜；这些物质的逆行运输是由内吞囊泡的裂变提供的。由于细胞膜的电特性由选择性电导和电容组成，后者与膜表面积成正比，因此可以通过记录膜电容直接监测囊泡融合和裂变引起的表面积变化。通过这种方式，可以在活细胞中实时记录单个外吞和内吞囊泡的大小及其融合和裂变动力学。在证明这种记录技术在酵母原生质体中也是可行的之后，我们将在本提案中将酵母中外吞/内吞作用的详细遗传和生化知识与高分辨率电容记录结合起来。使用在外吞和内吞机制中具有不同蛋白质缺陷的丰富的酵母突变体库，以及使用已建立的膜运输信号（如交配信息素）将使得将融合和裂变的复杂机制剖析成为可能。定义的分子步骤。由于分子机制的保守性，这项工作的结果将对理解哺乳动物细胞中的这些过程产生重大影响。