Isolation of quiescent and nonquiescent cells from yeast stationary-phase cultures

Chris Allen, Sabrina Büttner, Anthony D. Aragon, Jason A. Thomas, Osorio Meirelles, Jason E. Jaetao, Don Benn, Stephanie W. Ruby, Marten Veenhuis, Frank Madeo, Margaret Werner-Washburne*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Quiescence is the most common and, arguably, most poorly understood cell cycle state. This is in part because pure populations of quiescent cells are typically difficult to isolate. We report the isolation and characterization of quiescent and nonquiescent cells from stationary-phase (SP) yeast cultures by density-gradient centrifugation. Quiescent cells are dense, unbudded daughter cells formed after glucose exhaustion. They synchronously reenter the mitotic cell cycle, suggesting that they are in a G0 state. Nonquiescent cells are less dense, heterogeneous, and composed of replicatively older, asynchronous cells that rapidly lose the ability to reproduce. Microscopic and flow cytometric analysis revealed that nonquiescent cells accumulate more reactive oxygen species than quiescent cells, and over 21 d, about half exhibit signs of apoptosis and necrosis. The ability to isolate both quiescent and nonquiescent yeast cells from SP cultures provides a novel, tractable experimental system for studies of quiescence, chronological and replicative aging, apoptosis, and the cell cycle.
Original languageEnglish
Pages (from-to)89 - 100
Number of pages12
JournalJournal of Cell Biology
Volume174
Issue number1
DOIs
Publication statusPublished - 2006

Keywords

  • SACCHAROMYCES-CEREVISIAE
  • GENE-EXPRESSION
  • APOPTOSIS
  • STRESS
  • EXIT
  • DNA
  • IDENTIFICATION
  • SUPEROXIDE
  • REGULATOR
  • COLONIES

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