Proteomic analysis of the S. cerevisiae cell cycle

酿酒酵母细胞周期的蛋白质组学分析

• 批准号: 6587505
• 负责人: SINA GHAEMMAGHAMI
• 金额: $ 4.16万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-18 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6587505
• 关键词: Saccharomyces cerevisiae; affinity chromatography; cell cycle; cell cycle proteins; gene expression; high throughput technology; open reading frames; peptide library; postdoctoral investigator; posttranslational modifications; protein structure; proteomics

DESCRIPTION (provided by applicant): The coordinated transition between the stages of the eukaryotic cell cycle is controlled by a number of regulatory mechanisms at both the transcriptional and post-translational levels Errors in the regulation of the cell cycle lead to genetic instability and play a critical role in the onset and progression of many cancers This proposal investigates the post-translational regulation of cell cycle related proteins in S cerevisiae by utilizing a proteomics approach A fusion library will be created in which all known ORFs me tagged with a common epitope while keeping their expression under the control of their natural promoters The library will allow for endogenous protein levels to be quantified through the immunodetection of the fused tag. High throughput protocols will be developed in order to detect changes in the levels of all expressed proteins, as a function of the cell cycle, in 96-well format. This analysis will be used to identify, classify and study new cell cycle related proteins and to discover novel post-translational regulatory mechanisms. More detailed follow-up studies will provide further clues as to the exact role of the identified proteins. Also, the correlation between changes in mRNA and protein levels during the cell cycle will be investigated. The global-scale analysis of post-translational regulatory mechanisms will add to our understanding of cell cycle regulation and allow for the identification of novel genes that have relevance to the study of cancer More generally, it is hoped that the fusion library-based proteomic approach, employed here in the study the cell cycle, will in future serve as a general model for the investigation of other complex biological systems

描述（由申请人提供）： The coordinated transition between the stages of the eukaryotic cell cycle is controlled by a number of regulatory mechanisms at both the transcriptional and post-translational levels Errors in the regulation of the cell cycle lead to genetic instability and play a critical role in the onset and progression of many cancers This proposal investigates the post-translational regulation of cell cycle related proteins in S cerevisiae by utilizing a proteomics approach A将创建融合文库，其中所有已知的ORF我都用公共表位标记，同时将其表达在其天然启动子的控制下，库将通过融合标签的免疫检测来量化内源性蛋白质水平。将开发高吞吐量方案，以检测所有表达蛋白水平的变化，作为细胞周期的函数，以96孔格式。该分析将用于识别，分类和研究与细胞周期相关的新蛋白质，并发现新颖的翻译后调节机制。 更详细的随访研究将为确定的蛋白质的确切作用提供进一步的线索。同样，将研究mRNA和蛋白质水平的变化在细胞周期中的相关性。对翻译后调节机制的全球规模分析将增加我们对细胞周期调节的理解，并允许鉴定与癌症研究相关的新型基因，希望基于融合库的蛋白质组学方法，在研究中采用的研究细胞周期将来可以作为对其他复杂生物系统的一般模型的研究。