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 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中得到体现。列出的机构是中心的机构，不一定是研究者的机构。 Yra1p 及其脊椎动物同源物与 mRNA 输出因子 Mex67p/TAP 结合，被认为在体内 mRNA 输出中发挥作用。为了进一步表征 Yra1p，我们使用免疫亲和层析来纯化内源 Yra1p 复合物。这些实验表明，两个输入蛋白 β 同源物（Kap123p 和 Pse1p）以及聚 A 尾结合蛋白 Pab1p 和 Nab2p 与 Yra1p 相关。与 Yra1p 相关的其他主要蛋白质包括参与 mRNA 和 rRNA 加工的蛋白质以及 Yra1p 相关蛋白质 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。我们生成了 YRA1 的温度敏感等位基因，并用它来证明缺乏 Yra1p 和 Yra2p 功能的细胞能够退出 G0 停滞，并在停滞前经历几轮细胞分裂。我们还鉴定了 yra1-2 温度敏感生长缺陷的高拷贝抑制因子。其中包括 SUB2（一种在 mRNA 输出中重要的剪接因子）、ULP1（一种位于核孔并参与 Smt3p/SUMO 加工的核半胱氨酸蛋白酶）和 YRA2。总而言之，这些结果表明 Yra1p 除了在 mRNA 输出中发挥作用之外，还在多种 RNA 加工事件中发挥作用。