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.

虽然分泌途径是生物生存所必需的 对于单个细胞和整个有机体来说， 对囊泡运输的调控知之甚少。最近的数据显示 研究表明，ras相关的小gtp结合蛋白的rab家族是 参与调节不同物种之间的膜交通 亚细胞隔间。这一奖学金的长期目标是 建议是表征Rab 1基因产物在 蛋白质从内质网(ER)输出到高尔基复合体。 只有通过对正常细胞功能的了解，才能 可以开始了解缺陷在哪里可能导致疾病过程。 Ras蛋白的结构信息使其成为 有可能在ras初级序列中定位功能结构域。 Rab-1基因类似已知功能的定点突变 将生成RAS的域。这些Rab的突变形式(或者 在质粒或重组痘苗病毒中编码)将被 将其导入组织培养细胞，然后评估其对 体内的蛋白质交易。此外，体外转运试验 在这个实验室中开发的，它重新构建了 将使用Er和cis Golgi隔室来研究生化 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，是 分泌途径。