RAS-RELATED RABI AND VESICULAR TRANSPORT

RAS 相关的 RABI 和囊泡运输

• 批准号: 2085166
• 负责人: ELLEN J TISDALE
• 金额: $ 3.25万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-03-15 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2085166
• 关键词: Golgi apparatus; SDS polyacrylamide gel electrophoresis; Vesiculovirus; complementary DNA; endoplasmic reticulum; gene mutation; guanine nucleotide binding protein; hydrolysis; immunofluorescence technique; intracellular transport; molecular cloning; molecular genetics; monoclonal antibody; point mutation; posttranslational modifications; protein structure function; protein transport; recombinant DNA; site directed mutagenesis; tissue /cell culture; transfection; vaccinia virus

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.

尽管需要分泌途径才能生存 单个细胞和整个生物体的机制 囊泡运输的理解很少。 最近的数据有 表明与RAS相关的小型GTP结合蛋白的Rab家族是 参与不同的膜流量调节 亚细胞室。 该奖学金的长期目标 建议是表征Rab 1基因产物在 蛋白质从内质网（ER）出口到高尔基体复合物。 只有通过对正常细胞功能的知识才能 可以开始了解缺陷可能导致疾病过程的地方。 可用于RAS蛋白的结构信息已成为 可以在RAS主序列中绘制功能域。 Rab 1基因中的位置定向突变类似于已知功能 将生成RA的域。 这些突变形式的rab（要么 在质粒或重组牛ac病毒中编码）将是 引入组织培养细胞，然后评估它们对 蛋白质运输，体内。 另外，体外运输测定法 在这个实验室中开发的，该实验室重构是在 ER和顺式高尔基室将用于研究生化 Rab 1的功能并识别和隔离上游或下游 效应子分子与RAB 1相互作用，在 分泌道路。