Abstract
IntroductionThe switch from quiescence (G0) into G1 and cell cycle progression critically depends on specific nutrients and metabolic capabilities. Conversely, metabolic networks are regulated by enzyme–metabolite interaction and transcriptional regulation that lead to flux modifications to support cell growth. How cells process and integrate environmental information into coordinated responses is challenging to analyse and not yet described quantitatively.ObjectivesTo quantitatively monitor the central carbon metabolism during G0 exit and the first 2 h after reentering the cell cycle from synchronized Saccharomyces cerevisiae.MethodsDynamic tailored 13C metabolic flux analysis was used to observe the intracellular metabolite flux changes, and the metabolome and proteome were observed to identify regulatory mechanisms.ResultsG0 cells responded immediately to an extracellular increase of glucose. The intracellular metabolic flux changed in time and specific events were observed. High fluxes into trehalose and glycogen synthesis were observed during the G0 exit. Both fluxes then decreased, reaching a minimum at t = 65 min. Here, storage degradation contributed significantly (i.e. 21%) to the glycolytic flux. In contrast to these changes, the glucose uptake rate remained constant after the G0 exit. The flux into the oxidative pentose phosphate pathway was highest (29-fold increase, 36.4% of the glucose uptake) at t = 65 min, while it was very low at other time points. The maximum flux seems to correlate with a late G1 state preparing for the S phase transition. In the G1/S phase (t = 87 min), anaplerotic reactions such as glyoxylate shunt increased. Protein results show that during this transition, proteins belonging to clusters related with ribosome biogenesis and assembly, and initiation transcription factors clusters were continuously synthetised.ConclusionThe intracellular flux distribution changes dynamically and these major rearrangements highlight the coordinate reorganization of metabolic flux to meet requirements for growth during different cell state.
Highlights
The switch from quiescence (G0) into G1 and cell cycle progression critically depends on specific nutrients and metabolic capabilities
A S. cerevisiae culture was grown in mineral media without yeast extract and driven into G0 state by carbon starvation for 10 h (Tu et al 2005)
Of the population was in G0 and 11% was in G1
Summary
The switch from quiescence (G0) into G1 and cell cycle progression critically depends on specific nutrients and metabolic capabilities. How cells process and integrate environmental information into coordinated responses is challenging to analyse and not yet described quantitatively. Storage degradation contributed significantly (i.e. 21%) to the glycolytic flux In contrast to these changes, the glucose uptake rate remained constant after the G0 exit. Conclusion The intracellular flux distribution changes dynamically and these major rearrangements highlight the coordinate reorganization of metabolic flux to meet requirements for growth during different cell state. How cells process and integrate environmental information into coordinated physiological responses is still one of the most fundamental challenges in biology that is challenging to analyse and not yet described quantitatively (Ewald 2018). The detailed metabolic activity, i.e. the actual rate of metabolic reactions and pathways during the quiescence exit and subsequent cell cycle progression remain largely unknown
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