The proteomics of quiescent and nonquiescent cell differentiation in yeast stationary-phase cultures

• Published in: Molecular biology of the cell Vol. 22; no. 7; pp. 988 - 998
• Main Authors: Davidson, George S., Joe, Ray M., Roy, Sushmita, Meirelles, Osorio, Allen, Chris P., Wilson, Melissa R., Tapia, Phillip H., Manzanilla, Elaine E., Dodson, Anne E., Chakraborty, Swagata, Carter, Mark, Young, Susan, Edwards, Bruce, Sklar, Larry, Werner-Washburne, Margaret
• Format: Journal Article
• Language: English
• Published: United States American Society for Cell Biology 01.04.2011; The American Society for Cell Biology
• Subjects: BASIC BIOLOGICAL SCIENCES; Cell Biology; Cell Cycle - physiology; Flow Cytometry; Gene Expression Regulation, Fungal; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Oxygen Consumption; Proteomics; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Saccharomyces cerevisiae - cytology; Saccharomyces cerevisiae - physiology; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism
• ISSN: 1059-1524; 1939-4586; 1939-4586
• DOI: 10.1091/mbc.e10-06-0499

As yeast cultures enter stationary phase in rich, glucose-based medium, differentiation of two major subpopulations of cells, termed quiescent and nonquiescent, is observed. Differences in mRNA abundance between exponentially growing and stationary-phase cultures and quiescent and nonquiescent cells are known, but little was known about protein abundance in these cells. To measure protein abundance in exponential and stationary-phase cultures, the yeast GFP-fusion library (4159 strains) was examined during exponential and stationary phases, using high-throughput flow cytometry (HyperCyt). Approximately 5% of proteins in the library showed twofold or greater changes in median fluorescence intensity (abundance) between the two conditions. We examined 38 strains exhibiting two distinct fluorescence-intensity peaks in stationary phase and determined that the two fluorescence peaks distinguished quiescent and nonquiescent cells, the two major subpopulations of cells in stationary-phase cultures. GFP-fusion proteins in this group were more abundant in quiescent cells, and half were involved in mitochondrial function, consistent with the sixfold increase in respiration observed in quiescent cells and the relative absence of Cit1p:GFP in nonquiescent cells. Finally, examination of quiescent cell–specific GFP-fusion proteins revealed symmetry in protein accumulation in dividing quiescent and nonquiescent cells after glucose exhaustion, leading to a new model for the differentiation of these cells.