D'Urso G, Nurse P
University of Miami School of Medicine, Department of Biochemistry and Molecular Biology, Miami, FL 33136, USA.
Proc Natl Acad Sci U S A. 1997 Nov 11;94(23):12491-6. doi: 10.1073/pnas.94.23.12491.
In fission yeast both DNA polymerase alpha (pol alpha) and delta (pol delta) are required for DNA chromosomal replication. Here we demonstrate that Schizosaccharomyces pombe cdc20+ encodes the catalytic subunit of DNA polymerase epsilon (pol epsilon) and that this enzyme is also required for DNA replication. Following a shift to the restrictive temperature, cdc20 temperature-sensitive mutant cells block at the onset of DNA replication, suggesting that cdc20+ is required early in S phase very near to the initiation step. In the budding yeast Saccharomyces cerevisiae, it has been reported that in addition to its proposed role in chromosomal replication, DNA pol epsilon (encoded by POL2) also functions directly as an S phase checkpoint sensor [Navas, T. A., Zhou, Z. & Elledge, S. J. (1995) Cell 80, 29-39]. We have investigated whether cdc20+ is required for the checkpoint control operating in fission yeast, and our data indicate that pol epsilon does not have a role as a checkpoint sensor coordinating S phase with mitosis. In contrast, germinating spores disrupted for the gene encoding pol alpha rapidly enter mitosis in the absence of DNA synthesis, suggesting that in the absence of pol alpha, normal coordination between S phase and mitosis is lost. We propose that the checkpoint signal operating in S phase depends on assembly of the replication initiation complex, and that this signal is generated prior to the elongation stage of DNA synthesis.
在裂殖酵母中,DNA染色体复制需要DNA聚合酶α(polα)和δ(polδ)。在此我们证明,粟酒裂殖酵母cdc20⁺编码DNA聚合酶ε(polε)的催化亚基,并且这种酶对于DNA复制也是必需的。转移至限制温度后,cdc20温度敏感突变细胞在DNA复制开始时受阻,这表明cdc20⁺在S期早期靠近起始步骤时是必需的。在芽殖酵母酿酒酵母中,据报道,除了其在染色体复制中的假定作用外,DNA polε(由POL2编码)还直接作为S期检查点传感器发挥作用[Navas, T. A., Zhou, Z. & Elledge, S. J. (1995) Cell 80, 29 - 39]。我们研究了cdc20⁺对于裂殖酵母中运行的检查点控制是否必需,我们的数据表明polε不作为协调S期与有丝分裂的检查点传感器发挥作用。相反,编码polα的基因被破坏的萌发孢子在没有DNA合成的情况下迅速进入有丝分裂,这表明在没有polα的情况下,S期与有丝分裂之间的正常协调丧失。我们提出,在S期运行的检查点信号取决于复制起始复合物的组装,并且该信号在DNA合成的延伸阶段之前产生。