期刊
SYMMETRY-BASEL
卷 15, 期 9, 页码 -出版社
MDPI
DOI: 10.3390/sym15091793
关键词
Saccharomyces cerevisiae; cyclic AMP; FRET; protein kinase A; oscillations
This study reveals that the Ras/cAMP/PKA pathway plays a crucial role in regulating the response to nutrients' availability and stress in budding yeast. The activity of this pathway leads to noisy oscillations of cAMP levels, which in turn affects the nuclear accumulation of the transcription factor Msn2. Using a sensor, the researchers were able to detect the changes in cAMP concentrations in single yeast cells and observed oscillations under conditions of nutritional stress caused by limiting glucose availability.
The Ras/cAMP/PKA pathway regulates responses to nutrients' availability and stress in budding yeast. The cAMP levels are subjected to negative feedback, and we have previously simulated a dynamic model of this pathway suggesting the existence of stable oscillatory states depending on the symmetrical and opposed activity of the RasGEF (Cdc25) and RasGAPs (Ira proteins). Noisy oscillations related to the activity of this pathway were reported by looking at the nuclear localization of the transcription factor Msn2, and sustained oscillations of the nuclear accumulation of Msn2 under the condition of limiting glucose were observed. We were able to reproduce the periodic accumulation of Msn2-GFP protein in a yeast cell under the condition of limiting glucose, and we also detected oscillations of cAMP. We used a sensor based on a fusion protein between YFP-Epac2-CFP expressed in yeast cells. The FRET between CFP and YFP is controlled by the cAMP concentration. This sensor allows us to monitor changes in cAMP concentrations in a single yeast cell over a long time. Using this method, we were able to detect noisy oscillations of cAMP levels in single yeast cells under conditions of nutritional stress caused by limiting glucose availability.
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