ATP analog-sensitive Pat1 protein kinase for synchronous fission yeast meiosis at physiological temperature

作者：Cipak Lubos; Hyppa Randy W; Smith Gerald R^*; Gregan Juraj

来源：Cell Cycle, 2012, 11(8): 1626-1633.

DOI：10.4161/cc.20052

• meiosis
• synchronization
• fission yeast
• analog-sensitive
• Pat1 kinase

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.

• 出版日期2012-4-15