Abstract
The yeast Saccharomyces cerevisiae has been an excellent model system for cell cycle studies. Many such studies require cells synchronized in some particular portion of the cell cycle. Here, methods are described for obtaining and examining synchronized cells as they pass through one or more rounds of the cell cycle. The methods are of two types. First, block-and-release methods, where cells are initially synchronized by blocking them at some particular cell cycle stage, then releasing them from the block under conditions suitable for growth, and taking samples at different times after the release, thereby obtaining samples representing different cell cycle stages. The second type of method is elutriation. Centrifugal elutriation can be used to obtain samples of uniformly sized cells, and because cell size is correlated with cell cycle stage, these cells are synchronized with respect to their position in the cycle. Because elutriation is a very different method from block- and-release, it is ideal as a second method of synchronization to ensure that results achieved by block-and-release are not artefactual. Here, block-and-release experiments with the mating pheromone alpha factor, and with the cdc15-2 mutation, are described in detail, as are some elutriation methods.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Breeden LL (1997). α-factor synchronization of budding yeast. In: Dunphy WG (ed), Methods in Enzymology, pp 332–341. Cell Cycle Control. New York: Academic Press.
Carter BL, Jagadish MN (1978). The relationship between cell size and cell division in the yeast Saccharomyces cerevisiae. Exp Cell Res 112: 15–24.
Cho RJ, Campbell MJ, Winzeler EA, Steinmetz L, Conway A, Wodicka L, Wolfsberg TG, Gabrielian AE, Landsman D, Lockhart DJ, Davis RW (1998). A genome-wide transcriptional analysis of the mitotic cell cycle. Molecular Cell 2: 65–73.
Fitch I, Dahmann C, Surana U, Amon A, Nasmyth K, Goetsch L, Byers B, Futcher B (1992). Characterization of four B-type cyclin genes of the budding yeast S. cerevisiae. Mol Biol Cell 3: 805–818.
Futcher AB (1993). Analysis of the cell cycle in S. cerevisiae. In: Fantes P, Brooks R (eds), The cell cycle: A Practical Approach, pp 69–92. Oxford: IRL Press at Oxford University Press.
Haase SB, Lew DJ (1997). Flow cytometric analysis of DNA content in budding yeast. In: Dunphy WG (ed), Methods in enzymology, cell cycle control, pp 322–332. New York: Academic Press.
Johnson LH, Johnson AL (1997). Elutriation of budding yeast. In: Dunphy WG (ed), Methods in enzymology, cell cycle control, pp 342–350. New York: Academic Press.
Johnston LH, White JH, Johnson AL, Lucchini G, Plevani P (1990). Expression of the yeast DNA primase gene, PRI1, is regulated within the mitotic cell cycle and in meiosis. Mol Gen Genet 22: 44–48.
MacKay VL, Welch SK, Insley MY, Manney TR, Holly J, Saari GC, Parker ML (1988). The Saccharmoyces cerevisiae BAR1 gene encodes an exported protein with homology to pepsin. Proc Natl Acad Sci USA 85: 55–59.
McCarroll RM, Fangman WL (1988). Time of replication of yeast centromeres and telomeres. Cell 54: 505–513.
Oehlen LJ, Cross FR (1994). G1 cyclins CLN1 and CLN2 repress the mating factor response pathway at Start in the yeast cell cycle. Genes Dev 1: 1058–1070.
Oehlen LJ, McKinney JD, Cross FR (1996). Ste12 and Mcm1 regulate cell cycle-dependent transcription of FAR1. Mol Cell Biol 16: 2830–2837.
Pringle JR, Adams AEM, Drubin DG, et al. (1991). Immunofluorescence methods for yeast. In: Guthrie C, Fink GR (eds), Guide to yeast genetics and molecular biology, pp 565–602. New York: Academic Press.
Sherman F (1991). Getting started with yeast. In: Guthrie C, Fink GR (eds), Guide to yeast genetics and molecular biology, pp 3–21. New York: Academic Press.
Spellman PT, Sherlock G, Zhang MQ, Iyer VR, Anders K, Eisen MB, Brown PO, Botstein D, Futcher B (1998). Comprehensive identification of cell cycle regulated genes of the yeast S. cerevisiae by microarray hybridization. Mol Biol Cell 9: 3273–3297.
Van Doorn J, Valkenburg JA, Scholte ME, Oehlen LJ, Van Driel R, Postma PW, Nanninga N, Van Dam K (1988). Changes in activities of several enzymes involved in carbohydrate metabolism during the cell cycle of Saccharomyces cerevisiae. J Bacteriol 170: 4808–4815.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Futcher, B. Cell cycle synchronization. Methods Cell Sci 21, 79–86 (1999). https://doi.org/10.1023/A:1009872403440
Issue Date:
DOI: https://doi.org/10.1023/A:1009872403440