The proteomics of quiescent and nonquiescent cell differentiation in yeast stationary-phase cultures
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...
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| Published in | Molecular biology of the cell Vol. 22; no. 7; pp. 988 - 998 |
|---|---|
| Main Authors | , , , , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
American Society for Cell Biology
01.04.2011
The American Society for Cell Biology |
| Subjects | |
| Online Access | Get full text |
| ISSN | 1059-1524 1939-4586 1939-4586 |
| DOI | 10.1091/mbc.e10-06-0499 |
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| Abstract | 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. |
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| AbstractList | 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.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. 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. Starved yeast cultures differentiate into quiescent (Q) and nonquiescent (NQ) cell fractions. The yeast GFP-fusion library (4159 strains) and high-throughput flow cytometry were used to study this process. This showed significant metabolic and physiologic differences between Q/NQ cells and provided new tools for studying their differentiation. 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. |
| Author | Joe, Ray M. Young, Susan Tapia, Phillip H. Sklar, Larry Davidson, George S. Chakraborty, Swagata Dodson, Anne E. Wilson, Melissa R. Allen, Chris P. Edwards, Bruce Carter, Mark Werner-Washburne, Margaret Roy, Sushmita Manzanilla, Elaine E. Meirelles, Osorio |
| Author_xml | – sequence: 1 givenname: George S. surname: Davidson fullname: Davidson, George S. organization: Biology Department, University of New Mexico, Albuquerque, NM 87131, Sandia National Laboratories, Albuquerque, NM 81185 – sequence: 2 givenname: Ray M. surname: Joe fullname: Joe, Ray M. organization: Biology Department, University of New Mexico, Albuquerque, NM 87131 – sequence: 3 givenname: Sushmita surname: Roy fullname: Roy, Sushmita organization: Computer Science Department, University of New Mexico, Albuquerque, NM 87131 – sequence: 4 givenname: Osorio surname: Meirelles fullname: Meirelles, Osorio organization: Biology Department, University of New Mexico, Albuquerque, NM 87131 – sequence: 5 givenname: Chris P. surname: Allen fullname: Allen, Chris P. organization: Department of Cytometry, University of New Mexico, Albuquerque, NM 87131 – sequence: 6 givenname: Melissa R. surname: Wilson fullname: Wilson, Melissa R. organization: Biology Department, University of New Mexico, Albuquerque, NM 87131 – sequence: 7 givenname: Phillip H. surname: Tapia fullname: Tapia, Phillip H. organization: Department of Cytometry, University of New Mexico, Albuquerque, NM 87131 – sequence: 8 givenname: Elaine E. surname: Manzanilla fullname: Manzanilla, Elaine E. organization: Biology Department, University of New Mexico, Albuquerque, NM 87131 – sequence: 9 givenname: Anne E. surname: Dodson fullname: Dodson, Anne E. organization: Biology Department, University of New Mexico, Albuquerque, NM 87131 – sequence: 10 givenname: Swagata surname: Chakraborty fullname: Chakraborty, Swagata organization: Biology Department, University of New Mexico, Albuquerque, NM 87131 – sequence: 11 givenname: Mark surname: Carter fullname: Carter, Mark organization: Department of Cytometry, University of New Mexico, Albuquerque, NM 87131 – sequence: 12 givenname: Susan surname: Young fullname: Young, Susan organization: Department of Cytometry, University of New Mexico, Albuquerque, NM 87131 – sequence: 13 givenname: Bruce surname: Edwards fullname: Edwards, Bruce organization: Department of Cytometry, University of New Mexico, Albuquerque, NM 87131 – sequence: 14 givenname: Larry surname: Sklar fullname: Sklar, Larry organization: Department of Cytometry, University of New Mexico, Albuquerque, NM 87131, Department of Pathology, University of New Mexico School of Medicine, Albuquerque, NM 87131 – sequence: 15 givenname: Margaret surname: Werner-Washburne fullname: Werner-Washburne, Margaret organization: Biology Department, University of New Mexico, Albuquerque, NM 87131 |
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| Snippet | As yeast cultures enter stationary phase in rich, glucose-based medium, differentiation of two major subpopulations of cells, termed quiescent and... Starved yeast cultures differentiate into quiescent (Q) and nonquiescent (NQ) cell fractions. The yeast GFP-fusion library (4159 strains) and high-throughput... |
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| SubjectTerms | 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 |
| Title | The proteomics of quiescent and nonquiescent cell differentiation in yeast stationary-phase cultures |
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