Division of Systems and Synthetic Biology, Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden.
Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, Gothenburg, Sweden.
Yeast. 2021 Oct;38(10):549-565. doi: 10.1002/yea.3658. Epub 2021 Jul 1.
All living cells, including yeast cells, are challenged by different types of stresses in their environments and must cope with challenges such as heat, chemical stress, or oxidative damage. By reversibly adjusting the physiology while maintaining structural and genetic integrity, cells can achieve a competitive advantage and adapt environmental fluctuations. The yeast Saccharomyces cerevisiae has been extensively used as a model for study of stress responses due to the strong conservation of many essential cellular processes between yeast and human cells. We focused here on developing a tool to detect and quantify early responses using specific transcriptional responses. We analyzed the published transcriptional data on S. cerevisiae DBY strain responses to 10 different stresses in different time points. The principal component analysis (PCA) and the Pearson analysis were used to assess the stress response genes that are highly expressed in each individual stress condition. Except for these stress response genes, we also identified the reference genes in each stress condition, which would not be induced under stress condition and show stable transcriptional expression over time. We then tested our candidates experimentally in the CEN.PK strain. After data analysis, we identified two stress response genes (UBI4 and RRP) and two reference genes (MEX67 and SSY1) under heat shock (HS) condition. These genes were further verified by real-time PCR at mild (42°C), severe (46°C), to lethal temperature (50°C), respectively.
所有的活细胞，包括酵母细胞，在其环境中都会受到不同类型的压力的挑战，必须应对热、化学压力或氧化损伤等挑战。通过在保持结构和遗传完整性的同时可逆地调整生理机能，细胞可以获得竞争优势并适应环境波动。由于酵母和人类细胞之间许多重要细胞过程的强保守性，酵母 Saccharomyces cerevisiae 被广泛用作研究应激反应的模型。我们专注于开发一种使用特定转录反应来检测和量化早期反应的工具。我们分析了已发表的关于 S. cerevisiae DBY 菌株对 10 种不同应激在不同时间点的响应的转录数据。主成分分析（PCA）和 Pearson 分析用于评估在每种单独应激条件下高度表达的应激反应基因。除了这些应激反应基因，我们还在每种应激条件下确定了参考基因，这些基因在应激条件下不会被诱导，并且随时间表现出稳定的转录表达。然后，我们在 CEN.PK 菌株中通过实验测试了我们的候选基因。在数据分析后，我们在热激（HS）条件下确定了两个应激反应基因（UBI4 和 RRP）和两个参考基因（MEX67 和 SSY1）。通过实时 PCR 进一步在温和（42°C）、严重（46°C）和致死温度（50°C）下验证了这些基因。
