Lehmayer Leonie, Bernauer Lukas, Emmerstorfer-Augustin Anita
Institute of Molecular Biotechnology, Graz University of Technology, NAWI Graz, Petersgasse 14/II, 8010 Graz, Austria.
BioTechMed-Graz, 8010 Graz, Austria.
iScience. 2022 Aug 6;25(9):104888. doi: 10.1016/j.isci.2022.104888. eCollection 2022 Sep 16.
The auxin-inducible degron (AID) system is a useful technique to rapidly deplete any protein of interest "on-demand." In this study, we successfully established the AID system for the "biotech" yeast . First, we tested different expression levels of TIR1 for auxin-induced degradation of the glycerol kinase Gut1. Moderate expression of TIR1 resulted in complete degradation of the target protein within several minutes. Second, we show that the absence of all three Wsc type sensors is detrimental to cell growth, which indicates that these are the dominant cell wall sensors this yeast. Third, down-regulation of Erg1, an essential enzyme of the ergosterol biosynthetic pathway, resulted in quick and efficient accumulation of squalene, a pharmaceutically relevant reagent. We conclude that AID is an extremely powerful tool that, for the first time, enables the analysis of gene essentiality and function in .
生长素诱导降解标签（AID）系统是一种很有用的技术，可“按需”快速去除任何感兴趣的蛋白质。在本研究中，我们成功地为“生物技术”酵母建立了AID系统。首先，我们测试了不同表达水平的TIR1对生长素诱导甘油激酶Gut1降解的作用。TIR1的适度表达导致靶蛋白在几分钟内完全降解。其次，我们表明，所有三种Wsc型传感器的缺失对细胞生长有害，这表明它们是这种酵母的主要细胞壁传感器。第三，下调麦角固醇生物合成途径的关键酶Erg1，导致角鲨烯快速高效积累，角鲨烯是一种与药物相关的试剂。我们得出结论，AID是一种极其强大的工具，首次能够在……中分析基因的必需性和功能。
