ROLE OF RAS-RELATED RABI IN VESICULAR TRANSPORT

虽然分泌途径是生存所必需的， 对于单个细胞和整个生物体来说， 控制囊泡转运的机制知之甚少。 最近的数据 显示ras相关的小GTP结合蛋白的rab家族是 参与调节不同细胞之间的膜运输 亚细胞区室 本奖学金的长期目标是 建议是描述rab 1基因产物在 蛋白质从内质网（ER）输出到高尔基复合体。 只有通过了解正常细胞功能， 可以开始理解缺陷可能导致疾病的过程。 ras蛋白的结构信息使其 可以在ras一级序列中定位功能结构域。 rab 1基因中的定点突变类似于已知的功能性突变， 将产生RAS的结构域。 这些突变形式的rab（无论是 在质粒或重组牛痘病毒内编码）将被 引入到组织培养细胞中，然后评估它们对 蛋白质运输，在体内。 此外，体外转运试验 在这个实验室里开发的， ER和顺式高尔基体区室将被用来研究生物化学 rab 1的功能，并识别和隔离上游或下游 与rab 1相互作用的效应分子， 分泌途径

Although the secretory pathway is required for the survival of the individual cell and for the organism as a whole, the mechanism which governs vesicular transport is poorly understood. Recent data have shown that the rab family of ras-related small GTP-binding proteins are involved in the regulation of membrane traffic between distinct subcellular compartments. The long-term objective of this fellowship proposal is to characterize the role of the rab 1 gene product in protein export from the endoplasmic reticulum (ER) to the Golgi complex. It is only through the knowledge of normal cellular function that one can begin to understand where a defect could lead to a disease process. Structural information available for the ras protein has made it possible to map functional domains within the ras primary sequence. Site-directed mutations in the rab 1 gene analogous to known functional domains of ras will be generated. These mutated forms of rab (either encoded within a plasmid or a recombinant vaccinia virus) will be introduced into tissue culture cells, then evaluated for their effect on protein trafficking, in vivo. In addition, an in vitro transport assay developed in this laboratory which reconstitutes transport between the ER and cis Golgi compartments will be employed to study the biochemical function of rab 1 and to identify and isolate upstream or downstream effector molecules which interact with rab 1 and are required in the secretory pathway.