IDENTIFICATION OF NOVEL COMPONENTS IN TRANS-SNARE COMPLEX

这个子项目是利用资源的许多研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 膜融合是细胞生长（在正常细胞和癌细胞中）、激素分泌和神经传递的本质。 尽管它的基本特征，膜如何融合的机制只是慢慢地被理解。 我们用液泡来研究这个过程（溶酶体）从面包酵母的实验工具的无与伦比的力量可用：空泡很大，在活细胞中可以看到，它们的融合对于生存力是必需的或必需的。（取决于细胞生长条件），酵母的遗传学非常先进，我们可以用定量、比色测定法在体外测定液泡融合，并且融合反应已经用全纯化组分重构。 我们发现的机制，对于Rab-family GTdR，其效应复合物，以及SNARE，对于理解人类生理学和病理生理学具有普遍的重要性，因为这些相同的蛋白质对于人类的膜融合是必不可少的。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Membrane fusion is the essence of cell growth (in both normal cells and cancerous cells), hormone secretion, and neurotransmission. Despite its fundamental character, the mechanisms of how membranes fuse are only slowly becoming understood. We study this process with vacuoles (lysosomes) from baker's yeast for the unparalleled power of experimental tools available: vacuoles are large and can be visualized in the living cell, their fusion can be essential or dispensable for viability (depending on the cell growth conditions), the genetics of yeast is well-advanced, we can assay vacuole fusion in vitro with quantitative, colorimetric assays, and the fusion reaction has been reconstituted with all-purified components. The mechanisms we discover, for the Rab-family GTPase, for its effector complex, and for SNAREs, are of general importance for understanding human physiology and pathophysiology, for these same proteins are essential for membrane fusion in humans.