Improved flow cytometric analysis of the budding yeast cell cycle.
Steven B. Haase,Steven I. Reed +1 more
TLDR
The utility of the DNA binding dye, SYTOX Green, in the cell cycle analysis of yeast is shown, with better coefficients of variation, improved linearity between DNA content and fluorescence, and decreased peak drift associated with changes in dye concentration, growth conditions or cell size.Abstract:
The budding yeast, Saccharomyces cerevisiae has been a remarkably useful model system for the study of eukaryotic cell cycle regulation. Flow cytometric analysis of DNA content in budding yeast has become a standard tool for the analysis of cell cycle progression. However, popular protocols utilizing the DNA binding dye, propidium iodide, suffer from a number of drawbacks that confound accurate analysis by flow cytometry. Here we show the utility of the DNA binding dye, SYTOX Green, in the cell cycle analysis of yeast. Samples analyzed using SYTOX Green exhibited better coefficients of variation, improved linearity between DNA content and fluorescence, and decreased peak drift associated with changes in dye concentration, growth conditions or cell size.read more
Citations
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Cdc7-Dbf4 phosphorylates MCM proteins via a docking site-mediated mechanism to promote S phase progression.
Yi-Jun Sheu,Bruce Stillman +1 more
TL;DR: It is suggested that DDK docks on and phosphorylates MCM proteins at licensed origins to promote proper assembly of pre-IC, and genetic evidence suggests that phosphorylation of Mcm4 by DDK is important for timely S phase progression and for cell viability upon overproduction of Cdc45.
Prions are a common mechanism for phenotypic inheritance in wild yeasts
Randal Halfmann,Daniel F. Jarosz,Sandra K. Jones,Sandra K. Jones,Amelia N. Chang,Amelia N. Chang,Alex K. Lancaster,Susan Lindquist +7 more
TL;DR: Biochemically test approximately 700 wild strains of Saccharomyces for [PSI+] or [MOT3+], and find these prions in many, and they conferred diverse phenotypes that were frequently beneficial under selective conditions.
Journal ArticleDOI
Prions are a common mechanism for phenotypic inheritance in wild yeasts
Randal Halfmann,Daniel F. Jarosz,Sandra K. Jones,Sandra K. Jones,Amelia N. Chang,Amelia N. Chang,Alex K. Lancaster,Susan Lindquist,Susan Lindquist +8 more
TL;DR: In this paper, the self-templating conformations of yeast prion proteins act as epigenetic elements of inheritance, and they can provide a mechanism for generating heritable phenotypic diversity that promotes survival in fluctuating environments and the evolution of new traits.
Journal ArticleDOI
Symmetry-Breaking Polarization Driven by a Cdc42p GEF-PAK Complex
TL;DR: This work shows that Bem1p promotes symmetry breaking by assembling a complex in which both a Cdc42p-directed guanine nucleotide exchange factor (GEF) and a CDC42p effector p21-activated kinase (PAK) associate with Bem2p, and provides mechanistic insight into an evolutionarily conserved pattern-forming positive-feedback pathway.
Journal ArticleDOI
A Mechanism for Cell-Cycle Regulation of MAP Kinase Signaling in a Yeast Differentiation Pathway
Shelly Catherine Strickfaden,Matthew J. Winters,Giora Ben-Ari,Rachel E. Lamson,Mike Tyers,Mike Tyers,Peter M. Pryciak +6 more
TL;DR: A mechanism and physiological benefit of restricting antiproliferative signaling to G1 is defined, which suggests that Ste5 acts as a sensor for high G1 CDK activity and is an integration point for both external and internal signals.
References
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Journal ArticleDOI
Cyclin-dependent kinases prevent DNA re-replication through multiple mechanisms.
TL;DR: It is shown that B-type CDKs in Saccharomyces cerevisiae prevent re-initiation through multiple overlapping mechanisms, including phosphorylation of the origin recognition complex (ORC), downregulation of Cdc6 activity, and nuclear exclusion of the Mcm2-7 complex.
Journal ArticleDOI
Altered Fidelity of Mitotic Chromosome Transmission in Cell Cycle Mutants of S. CEREVISIAE
TL;DR: It is suggested that special aspects of DNA metabolism may be occurring in G2 and M in order to prepare the chromosomes for proper segregation and a delay in almost any stage of chromosome replication or segregation leads to a decrease in the fidelity of mitotic chromosome transmission.
Journal ArticleDOI
Characterization of four B-type cyclin genes of the budding yeast Saccharomyces cerevisiae.
I. Fitch,Christian Dahmann,Uttam Surana,Angelika Amon,Kim Nasmyth,L Goetsch,B Byers,Bruce Futcher +7 more
TL;DR: The sequences of another related pair of B-type cyclin genes, which are term CLB3 and CLB4, are presented and it is suggested that the two groups of Clbs may have distinct roles in spindle formation and elongation.
Journal ArticleDOI
Cyclin-B homologs in Saccharomyces cerevisiae function in S phase and in G2.
TL;DR: Cloned four cyclin-B homologs from Saccharomyces cerevisiae, CLB1-CLB4, show that the CLB genes perform an essential role at the G2/M-phase transition, and also a role in S phase, while CLB3 and CLB4 might participate additionally in DNA replication and spindle assembly.
Journal ArticleDOI
Cdc53p acts in concert with cdc4p and cdc34p to control the g1-to-s-phase transition and identifies a conserved family of proteins
Neal Mathias,Stephen L. Johnson,Stephen L. Johnson,Mark Winey,Mark Winey,Alison E.M. Adams,Alison E.M. Adams,Loretta Goetsch,John R. Pringle,Breck Byers,Mark G. Goebl +10 more
TL;DR: Evidence indicating that CDC53 also is involved in the targeted degradation of both activating and inhibitory subunits of the cyclin-dependent kinases is described, suggesting a role for this family of proteins in regulating cell cycle proliferation through protein degradation.
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