ISOLATION AND ANALYSIS OF PROTEINS WHICH REGULATE RAS

调节 RAS 的蛋白质的分离和分析

• 批准号: 3194657
• 负责人: DANIEL L BROEK
• 金额: $ 16.78万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 1994-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3194657
• 关键词: Saccharomyces cerevisiae; biochemical evolution; biological signal transduction; cell cycle; cell cycle proteins; cell growth regulation; chimeric proteins; complementary DNA; cytogenetics; drug design /synthesis /production; enzyme mechanism; fungal genetics; gel electrophoresis; gene expression; gene mutation; genetic manipulation; genetic regulation; guanine nucleotide binding protein; human genetic material tag; human tissue; immunological substance; laboratory rabbit; molecular genetics; mutagen testing; nucleic acid probes; nucleic acid sequence; oncogenes; posttranslational modifications; protein purification; protein structure function; radionuclides; reagent /indicator; regulatory gene; transcription factor

The regulation of cell division in mammalian cells is a complex process; coordinating growth promoting signals transduced across the cytoplasmic membrane initiated by growth factors, with signals initiated by elements monitoring the nutritional status of the cells. The discovery of cellular oncogenes provide a means to study, at least in part, the biochemical mechanisms controlling cell division. One group of these oncogenes, the ras genes, are found to be mutationally activated in a large number and wide variety of human tumors. The ras genes are highly conserved in evolution, suggesting they play a fundamental role in the biology of the cell. Both the function of the ras proteins and how the ras proteins are regulated remains a mystery for mammalian cells. The discovery of the yeast RAS genes made possible a systematic approach to study RAS protein function because of the genetic manipulation possible in that organism. This proposal is aimed at studying the CDC25 proteins in yeast which can regulate, not only yeast RAS protein function but the human H-ras protein as well. This regulation is likely due to a direct protein-protein interaction between CDC25 and RAS proteins. The study of this protein-protein interaction and its conservation t rough evolution provides a unique means to identify the mammalian molecule which regulates ras protein function. We will develop DNA probes and immunological reagents which encode or recognize the domain of the CDC25 protein essential for RAS activation. These reagents will be used to clone cDNAs encoding domains homologous to CDC25. Further, we will use a genetic screen to identify human cDNAs which can complement loss of CDC25 function in yeast. The identification of the molecular mechanism of action of the human ras proteins may allow the design of molecularly rational drugs to reverse the tumorigenic effect of mutant ras proteins.

哺乳动物细胞的细胞分裂调控是一个复杂的过程；