Guttmann-Raviv N, Boger-Nadjar E, Edri I, Kassir Y
Department of Biology, Technion, Technion City, Haifa 32000, Israel.
Genetics. 2001 Dec;159(4):1547-58. doi: 10.1093/genetics/159.4.1547.
In the budding yeast Saccharomyces cerevisiae initiation and progression through the mitotic cell cycle are determined by the sequential activity of the cyclin-dependent kinase Cdc28. The role of this kinase in entry and progression through the meiotic cycle is unclear, since all cdc28 temperature-sensitive alleles are leaky for meiosis. We used a "heat-inducible Degron system" to construct a diploid strain homozygous for a temperature-degradable cdc28-deg allele. We show that this allele is nonleaky, giving no asci at the nonpermissive temperature. We also show, using this allele, that Cdc28 is not required for premeiotic DNA replication and commitment to meiotic recombination. IME2 encodes a meiosis-specific hCDK2 homolog that is required for the correct timing of premeiotic DNA replication, nuclear divisions, and asci formation. Moreover, in ime2Delta diploids additional rounds of DNA replication and nuclear divisions are observed. We show that the delayed premeiotic DNA replication observed in ime2Delta diploids depends on a functional Cdc28. Ime2Delta cdc28-4 diploids arrest prior to initiation of premeiotic DNA replication and meiotic recombination. Ectopic overexpression of Clb1 at early meiotic times advances premeiotic DNA replication, meiotic recombination, and nuclear division, but the coupling between these events is lost. The role of Ime2 and Cdc28 in initiating the meiotic pathway is discussed.
在出芽酵母酿酒酵母中,有丝分裂细胞周期的起始和进程由细胞周期蛋白依赖性激酶Cdc28的顺序活性决定。该激酶在减数分裂周期的进入和进程中的作用尚不清楚,因为所有cdc28温度敏感等位基因在减数分裂方面都是渗漏型的。我们使用“热诱导降解子系统”构建了一个纯合温度可降解的cdc28-deg等位基因的二倍体菌株。我们表明该等位基因是无渗漏的,在非允许温度下不产生子囊。我们还使用该等位基因表明,减数分裂前DNA复制和减数分裂重组的起始不需要Cdc28。IME2编码一种减数分裂特异性的hCDK2同源物,它是减数分裂前DNA复制、核分裂和子囊形成正确时间所必需的。此外,在ime2Δ二倍体中观察到额外的DNA复制和核分裂轮次。我们表明在ime2Δ二倍体中观察到的减数分裂前DNA复制延迟取决于功能性的Cdc28。Ime2Δ cdc28-4二倍体在减数分裂前DNA复制和减数分裂重组起始之前停滞。在减数分裂早期异位过表达Clb1会促进减数分裂前DNA复制、减数分裂重组和核分裂,但这些事件之间的偶联会丧失。本文讨论了Ime2和Cdc28在启动减数分裂途径中的作用。