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粟酒裂殖酵母cdc20+编码DNA聚合酶ε,是染色体复制所必需的,但不是S期检查点所必需的。

Schizosaccharomyces pombe cdc20+ encodes DNA polymerase epsilon and is required for chromosomal replication but not for the S phase checkpoint.

作者信息

D'Urso G, Nurse P

机构信息

University of Miami School of Medicine, Department of Biochemistry and Molecular Biology, Miami, FL 33136, USA.

出版信息

Proc Natl Acad Sci U S A. 1997 Nov 11;94(23):12491-6. doi: 10.1073/pnas.94.23.12491.

DOI:10.1073/pnas.94.23.12491
PMID:9356477
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC25014/
Abstract

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.

摘要

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。

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Schizosaccharomyces pombe cdc20+ encodes DNA polymerase epsilon and is required for chromosomal replication but not for the S phase checkpoint.粟酒裂殖酵母cdc20+编码DNA聚合酶ε,是染色体复制所必需的,但不是S期检查点所必需的。
Proc Natl Acad Sci U S A. 1997 Nov 11;94(23):12491-6. doi: 10.1073/pnas.94.23.12491.
2
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本文引用的文献

1
Loading of an Mcm protein onto DNA replication origins is regulated by Cdc6p and CDKs.Mcm蛋白加载到DNA复制起点受Cdc6p和周期蛋白依赖性激酶(CDKs)调控。
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Cdc6p-dependent loading of Mcm proteins onto pre-replicative chromatin in budding yeast.芽殖酵母中Cdc6p介导的Mcm蛋白加载到复制前染色质上的过程。
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Cell cycle control of replication initiation in eukaryotes.真核生物中复制起始的细胞周期控制
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The ORC1 homolog orp1 in fission yeast plays a key role in regulating onset of S phase.裂殖酵母中的ORC1同源物orp1在调控S期起始过程中起关键作用。
Genes Dev. 1996 Oct 15;10(20):2644-54. doi: 10.1101/gad.10.20.2644.
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Cell cycle control of eukaryotic DNA replication.真核生物DNA复制的细胞周期调控。
Curr Opin Genet Dev. 1996 Apr;6(2):208-14. doi: 10.1016/s0959-437x(96)80052-9.
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Replication origins in eukaroytes.真核生物中的复制起点
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7
DNA polymerase epsilon may be dispensable for SV40- but not cellular-DNA replication.DNA聚合酶ε对于SV40介导的DNA复制可能并非必需,但对于细胞DNA复制却是必需的。
EMBO J. 1996 May 1;15(9):2298-305.
8
Identification of eukaryotic DNA replication proteins using simian virus 40 in vitro replication system.利用猿猴病毒40体外复制系统鉴定真核生物DNA复制蛋白
Methods Enzymol. 1995;262:522-48. doi: 10.1016/0076-6879(95)62043-5.
9
DNA polymerase alpha, a component of the replication initiation complex, is essential for the checkpoint coupling S phase to mitosis in fission yeast.DNA聚合酶α是复制起始复合物的一个组成部分,对于裂殖酵母中连接S期与有丝分裂的检查点至关重要。
J Cell Sci. 1995 Sep;108 ( Pt 9):3109-18. doi: 10.1242/jcs.108.9.3109.
10
Dpb11, which interacts with DNA polymerase II(epsilon) in Saccharomyces cerevisiae, has a dual role in S-phase progression and at a cell cycle checkpoint.在酿酒酵母中与DNA聚合酶II(ε)相互作用的Dpb11在S期进程和细胞周期检查点中具有双重作用。
Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11791-5. doi: 10.1073/pnas.92.25.11791.