title: ATP analog-sensitive Pat1 protein kinase for synchronous fission yeast meiosis at physiological temperature.
abstract: To study meiosis, synchronous cultures are often indispensable, especially for physical analyses of DNA and proteins. A temperature-sensitive allele of the Pat1 protein kinase (pat1-114) has been widely used to induce synchronous meiosis in the fission yeast Schizosaccharomyces pombe, but pat1-114-induced meiosis differs from wild-type meiosis, and some of these abnormalities might be due to higher temperature needed to inactivate the Pat1 kinase. Here, we report an ATP analog-sensitive allele of Pat1 [Pat1(L95A), designated pat1-as2] that can be used to generate synchronous meiotic cultures at physiological temperature. In pat1-as2 meiosis, chromosomes segregate with higher fidelity, and spore viability is higher than in pat1-114 meiosis, although recombination is lower by a factor of 2-3 in these mutants than in starvation-induced pat1(+) meiosis. Addition of the mat-Pc gene improved chromosome segregation and spore viability to nearly the level of starvation-induced meiosis. We conclude that pat1-as2 mat-Pc cells offer synchronous meiosis with most tested properties similar to those of wild-type meiosis.
authors.affiliation: Max F. Perutz Laboratories, University of Vienna, Vienna, Austria.
authors.forename: Lubos
authors.initials: L
authors.lastname: Cipak
authors.forename: Randy W
authors.initials: RW
authors.lastname: Hyppa
authors.forename: Gerald R
authors.initials: GR
authors.lastname: Smith
authors.forename: Juraj
authors.initials: J
authors.lastname: Gregan
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology*; D000483:Alleles; D020090:Chromosome Segregation / Q000187:drug effects; D008540:Meiosis / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012568:Schizosaccharomyces / Q000166:cytology / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D013172:Spores, Fungal / Q000187:drug effects; D013696:Temperature
chemical_list: D029702:Schizosaccharomyces pombe Proteins; D000255:Adenosine Triphosphate; C087019:ran1 protein, S pombe; D017346:Protein Serine-Threonine Kinases