Modeling single-cell phenotypes links yeast stress acclimation to transcriptional repression and pre-stress cellular states.
Modeling single-cell phenotypes links yeast stress acclimation to transcriptional repression and pre-stress cellular states.
复制标题
DOI：
10.7554/elife.82017
复制
发表时间：
2022-11-09
期刊：
影响因子：
7.7
通讯作者：
Gasch AP
中科院分区：
文献类型：
--
作者：
Bergen AC;Kocik RA;Hose J;McClean MN;Gasch AP
在大量培养分析中，应激防御和细胞生长呈负相关;然而，这些研究遗漏了大量的细胞间异质性，从而模糊了真实的表型关系。在这里，我们设计了一个微流体系统，以表征盐胁迫之前，期间和之后随着时间的推移在单个酵母细胞中的多种表型。该系统测量细胞和菌落大小，生长速率和细胞周期沿着核内两个转录因子的转定位：应激激活的Msn 2，调节防御基因和Dot 6，抑制核糖体生物合成基因在积极的应激反应。通过动态跟踪细胞，我们发现了Msn 2和Dot 6行为之间意想不到的不一致性，揭示了具有不同生长特性的细胞亚群。令人惊讶的是，应激后生长恢复与Dot 6阻遏物的激活呈正相关。相反，缺乏Dot 6的细胞显示出较慢的生长适应，即使它们在没有应激的情况下正常生长。我们发现，野生型细胞具有较大的Dot 6反应显示更快的生产Msn 2调节的Ctt 1蛋白，可与Msn 2的贡献。这些结果与酵母中急性应激期间的转录抑制提供保护性反应的模型一致，可能是通过将翻译能力重定向到诱导的转录物。
Stress defense and cell growth are inversely related in bulk culture analyses; however, these studies miss substantial cell-to-cell heterogeneity, thus obscuring true phenotypic relationships. Here, we devised a microfluidics system to characterize multiple phenotypes in single yeast cells over time before, during, and after salt stress. The system measured cell and colony size, growth rate, and cell-cycle phase along with nuclear trans-localization of two transcription factors: stress-activated Msn2 that regulates defense genes and Dot6 that represses ribosome biogenesis genes during an active stress response. By tracking cells dynamically, we discovered unexpected discordance between Msn2 and Dot6 behavior that revealed subpopulations of cells with distinct growth properties. Surprisingly, post-stress growth recovery was positively corelated with activation of the Dot6 repressor. In contrast, cells lacking Dot6 displayed slower growth acclimation, even though they grow normally in the absence of stress. We show that wild-type cells with a larger Dot6 response display faster production of Msn2-regulated Ctt1 protein, separable from the contribution of Msn2. These results are consistent with the model that transcriptional repression during acute stress in yeast provides a protective response, likely by redirecting translational capacity to induced transcripts.