Extreme calorie restriction in yeast retentostats induces uniform non-quiescent growth arrest.
Non-dividing Saccharomyces cerevisiae cultures are highly relevant for fundamental and applied studies. However, cultivation conditions in which non-dividing cells retain substantial metabolic activity are lacking. Unlike stationary-phase (SP) batch cultures, the current experimental paradigm for non-dividing yeast cultures, cultivation under extreme calorie restriction (ECR) in retentostat enables non-dividing yeast cells to retain substantial metabolic activity and to prevent rapid cellular deterioration. Distribution of F-actin structures and single-cell copy numbers of specific transcripts revealed that cultivation under ECR yields highly homogeneous cultures, in contrast to SP cultures that differentiate into quiescent and non-quiescent subpopulations. Combined with previous physiological studies, these results indicate that yeast cells subjected to ECR survive in an extended G1phase. This study demonstrates that yeast cells exposed to ECR differ from carbon-starved cells and offer a promising experimental model for studying non-dividing, metabolically active, and robust eukaryotic cells.
Department of Biotechnology, Delft University of Technology, van der Maasweg 9, 2629 HZ Delft, The Netherlands.
Markus M M
MM
Bisschops
Department of Biotechnology, Delft University of Technology, van der Maasweg 9, 2629 HZ Delft, The Netherlands.
Marijke A H
MA
Luttik
Systems Bioinformatics, VU University Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands.
Anne
A
Doerr
Department of Biotechnology, Delft University of Technology, van der Maasweg 9, 2629 HZ Delft, The Netherlands.
Peter J T
PJ
Verheijen
Systems Bioinformatics, VU University Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands.
Frank
F
Bruggeman
Department of Biotechnology, Delft University of Technology, van der Maasweg 9, 2629 HZ Delft, The Netherlands.
Jack T
JT
Pronk
Department of Biotechnology, Delft University of Technology, van der Maasweg 9, 2629 HZ Delft, The Netherlands. Electronic address: P.A.S.Daran-Lapujade@TUDelft.NL.
Pascale
P
Daran-Lapujade
D000199:Actins / Q000235:genetics / Q000378:metabolism; D061249:Batch Cell Culture Techniques; D019149:Bioreactors; D059447:Cell Cycle Checkpoints / Q000235:genetics*; D003470:Culture Media / Q000737:chemistry; D004734:Energy Metabolism / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D005947:Glucose / Q000172:deficiency*; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism
D000199:Actins; D003470:Culture Media; C112591:HSP12 protein, S cerevisiae; C499528:HSP26 protein, S cerevisiae; D006360:Heat-Shock Proteins; D029701:Saccharomyces cerevisiae Proteins; D005947:Glucose
Actin structure; Extreme calorie restriction; Heterogeneity; Non-dividing; Retentostat; mRNA FISH