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。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级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。尽管膜具有基本的特性，但膜如何融合的机制还只是慢慢地被了解。我们用面包酵母中的液泡(溶酶体)来研究这一过程，因为可用的实验工具具有无与伦比的能力：空泡很大，可以在活细胞中看到，它们的融合对生存能力来说可能是必不可少的，也可能是必不可少的(取决于细胞生长条件)，酵母的遗传学很先进，我们可以用定量、比色分析来检测体外的液泡融合，融合反应已经用全部纯化的成分重新组成。我们发现的Rab家族GTP酶、其效应复合体和SNARS的机制对于理解人类生理学和病理生理学具有普遍重要的意义，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.