CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
College of Life Sciences, University of Chinese Academy of Sciences, Beijing, China.
Appl Microbiol Biotechnol. 2023 Jul;107(14):4605-4619. doi: 10.1007/s00253-023-12593-2. Epub 2023 May 30.
Cat8 is a C6 zinc cluster transcription activator in yeast. It is generally recognized that the transcription of CAT8 is inhibited and that Cat8 is inactive in the presence of high concentrations of glucose. However, our recent study found that constitutively overexpressed Cat8 played a regulatory role in Saccharomyces cerevisiae in the presence of 20 g/L glucose. To explore the regulatory network of Cat8 at high glucose concentrations, CAT8 was both overexpressed and deleted in this study. Cell growth and glucose consumption in different media were significantly accelerated by the deletion of CAT8, while the lag period was greatly shortened. RNA-seq and genetic modification showed that the deletion of CAT8 changed the type of energy metabolism in yeast cells. Many genes related to the mitochondrial respiratory chain were downregulated, resulting in a reduction in aerobic respiration and the tricarboxylic acid cycle. Meanwhile, both the energy supply of anaerobic ethanol fermentation and the Crabtree effect of S. cerevisiae were enhanced by the deletion of CAT8. CAT8 knockout cells show a higher sugar uptake rate, a higher cell growth rate, and higher tolerance to glucose than the wild-type strain YS58. This study expands the understanding of the regulatory network of Cat8 and provides guidance for modulating yeast cell growth. KEY POINTS: • The deletion of CAT8 promoted cell growth of S. cerevisiae. • Transcriptome analysis revealed the regulation network of Cat8 under 1% glucose condition. • CAT8 deletion increases the glucose tolerance of cells by enhancing the Crabtree effect.
Cat8 是酵母中一种 C6 锌簇转录激活因子。一般认为,在高浓度葡萄糖存在的情况下,CAT8 的转录受到抑制,Cat8 处于无活性状态。然而,我们最近的研究发现,在 20 g/L 葡萄糖存在的情况下,组成性过表达的 Cat8 在酿酒酵母中发挥调节作用。为了探索高葡萄糖浓度下 Cat8 的调控网络,本研究同时过表达和敲除了 CAT8。CAT8 的缺失显著加速了不同培养基中细胞生长和葡萄糖消耗,大大缩短了迟滞期。RNA-seq 和遗传修饰表明,Cat8 的缺失改变了酵母细胞的能量代谢类型。许多与线粒体呼吸链相关的基因下调,导致有氧呼吸和三羧酸循环减少。同时,Cat8 的缺失增强了酵母细胞的无氧乙醇发酵的能量供应和 Crabtree 效应。Cat8 敲除细胞比野生型菌株 YS58 具有更高的糖摄取率、更高的细胞生长速率和更高的葡萄糖耐受性。本研究扩展了对 Cat8 调控网络的理解,并为调节酵母细胞生长提供了指导。 要点: • Cat8 的缺失促进了酿酒酵母的细胞生长。 • 转录组分析揭示了 1%葡萄糖条件下 Cat8 的调控网络。 • Cat8 缺失通过增强 Crabtree 效应提高了细胞的葡萄糖耐受性。