title: Synchronized fission yeast meiosis using an ATP analog-sensitive Pat1 protein kinase.
abstract: Synchronous cultures are often indispensable for studying meiosis. Here we present an optimized protocol for induction of synchronous meiosis in the fission yeast Schizosaccharomyces pombe. Chemical inactivation of an ATP analog-sensitive form of the Pat1 kinase (pat1-as2) by adding the ATP analog 1-NM-PP1 in G1-arrested cells allows the induction of synchronous meiosis at optimal temperature (25°C). Importantly, this protocol eliminates detrimental effects of elevated temperature (34°C), which is required to inactivate the commonly used temperature-sensitive Pat1 kinase mutant (pat1-114). The addition of the mat-Pc gene to a mat1-M strain further improves chromosome segregation and spore viability. Thus, our protocol offers highly synchronous meiosis at optimal temperature, with most characteristics similar to those of wild-type meiosis. The synchronization protocol can be completed in 5 d (not including strain production, which may take as long as 2 or 3 months).
authors.affiliation: 1] Max F. Perutz Laboratories, University of Vienna, Vienna, Austria. [2] Cancer Research Institute, Slovak Academy of Sciences, Bratislava, Slovakia. [3].
authors.forename: Lubos
authors.initials: L
authors.lastname: Cipak
authors.affiliation: 1] Max F. Perutz Laboratories, University of Vienna, Vienna, Austria. [2].
authors.forename: Silvia
authors.initials: S
authors.lastname: Polakova
authors.affiliation: 1] Fred Hutchinson Cancer Research Center, Division of Basic Sciences, Seattle, Washington, USA. [2].
authors.forename: Randy W
authors.initials: RW
authors.lastname: Hyppa
authors.affiliation: Fred Hutchinson Cancer Research Center, Division of Basic Sciences, Seattle, Washington, USA.
authors.forename: Gerald R
authors.initials: GR
authors.lastname: Smith
authors.affiliation: 1] Max F. Perutz Laboratories, University of Vienna, Vienna, Austria. [2] Department of Genetics, Faculty of Natural Sciences, Comenius University, Bratislava, Slovakia.
authors.forename: Juraj
authors.initials: J
authors.lastname: Gregan
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D018929:Cell Culture Techniques; D008540:Meiosis / Q000187:drug effects*; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000235:genetics; D012568:Schizosaccharomyces / Q000166:cytology*; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry* / Q000235:genetics; D013696:Temperature
chemical_list: D029702:Schizosaccharomyces pombe Proteins; D000255:Adenosine Triphosphate; C087019:ran1 protein, S pombe; D017346:Protein Serine-Threonine Kinases