BIOCHEMICAL AND GENETIC CHARACTERIZATION OF YRA1P IN BUDDING YEAST

芽殖酵母中 YRA1P 的生化和遗传特性

• 批准号: 7420661
• 负责人: Douglas R. Kellogg
• 金额: $ 0.29万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-20 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7420661
• 关键词: Chordata; RNA; alleles; binding proteins; cell cycle; cysteine; endopeptidases; immunoaffinity chromatography; molecular genetics; play; protein binding; proteins; role; temperature; thinking; yeasts

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Yra1p and its vertebrate homologues bind to the mRNA export factor Mex67p/TAP and are thought to play a role in mRNA export in vivo. To further characterize Yra1p, we used immunoaffinity chromatography to purify endogenous Yra1p complexes. These experiments demonstrated that two importin beta homologues (Kap123p and Pse1p) and the poly A tail-binding proteins Pab1p and Nab2p associate with Yra1p. The other major proteins that associate with Yra1p include proteins involved in mRNA and rRNA processing and the Yra1p-related protein Yra2p. Additional biochemical and genetic experiments suggest a close functional relationship between Yra1p and Yra2p. We generated a temperature-sensitive allele of YRA1 and used it to demonstrate that cells which lack the function of both Yra1p and Yra2p are able to exit a G0 arrest and go through several rounds of cell division before arresting. We also identified high-copy suppressors of the yra1-2 temperature-sensitive growth defect. These include SUB2, a splicing factor important in mRNA export, ULP1, a nuclear cysteine protease localized to the nuclear pore and involved in Smt3p/SUMO processing, and YRA2. Taken together, these results suggest that Yra1p has roles in diverse RNA processing events in addition to a role in mRNA export.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子弹和调查员（PI）可能已经从其他NIH来源获得了主要资金，因此可以在其他清晰的条目中代表。列出的机构适用于该中心，这不一定是调查员的机构。 YRA1P及其脊椎动物的同源物与mRNA输出因子MEX67P/TAP结合，并被认为在体内MRNA导出中起作用。为了进一步表征YRA1P，我们使用免疫亲和力色谱法纯化了内源性YRA1P复合物。这些实验表明，两个Importin beta同源物（KAP123P和PSE1P）和poly A尾结合蛋白PAB1P和NAB2P与YRA1P相关。与YRA1P关联的其他主要蛋白质包括参与mRNA和RRNA加工的蛋白质以及与YRA1P相关的蛋白YRA2P。其他生化和遗传实验表明YRA1P和YRA2P之间存在密切的功能关系。我们产生了YRA1温度敏感的等位基因，并用它来证明缺乏YRA1P和YRA2P功能的细胞能够退出G0停滞，并在逮捕之前经过几轮细胞分裂。我们还确定了YRA1-2温度敏感生长缺陷的高复制抑制剂。其中包括SUB2，这是一个在mRNA输出中重要的剪接因子，ULP1，一种局部在核孔中并参与SMT3P/SUMO加工的核半胱氨酸蛋白酶和YRA2。综上所述，这些结果表明YRA1P除了在mRNA输出中发挥作用外，在不同的RNA处理事件中还具有作用。