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

被引:73
作者
DUrso, G [1 ]
Nurse, P [1 ]
机构
[1] IMPERIAL CANC RES FUND, CELL CYCLE LAB, LONDON WC2A 3PX, ENGLAND
基金
英国惠康基金;
关键词
D O I
10.1073/pnas.94.23.12491
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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 po 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.
引用
收藏
页码:12491 / 12496
页数:6
相关论文
共 45 条
[1]   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 [J].
ARAKI, H ;
LEEM, SH ;
PHONGDARA, A ;
SUGINO, A .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (25) :11791-11795
[2]   DNA POLYMERASE-II, THE PROBABLE HOMOLOG OF MAMMALIAN DNA POLYMERASE-EPSILON, REPLICATES CHROMOSOMAL DNA IN THE YEAST SACCHAROMYCES-CEREVISIAE [J].
ARAKI, H ;
ROPP, PA ;
JOHNSON, AL ;
JOHNSTON, LH ;
MORRISON, A ;
SUGINO, A .
EMBO JOURNAL, 1992, 11 (02) :733-740
[3]   YEAST ORIGIN RECOGNITION COMPLEX FUNCTIONS IN TRANSCRIPTION SILENCING AND DNA-REPLICATION [J].
BELL, SP ;
KOBAYASHI, R ;
STILLMAN, B .
SCIENCE, 1993, 262 (5141) :1844-1849
[4]   ATP-DEPENDENT RECOGNITION OF EUKARYOTIC ORIGINS OF DNA-REPLICATION BY A MULTIPROTEIN COMPLEX [J].
BELL, SP ;
STILLMAN, B .
NATURE, 1992, 357 (6374) :128-134
[5]   A CONSERVED 3'-]5' EXONUCLEASE ACTIVE-SITE IN PROKARYOTIC AND EUKARYOTIC DNA-POLYMERASES [J].
BERNAD, A ;
BLANCO, L ;
LAZARO, JM ;
MARTIN, G ;
SALAS, M .
CELL, 1989, 59 (01) :219-228
[6]  
Brush GS, 1995, METHOD ENZYMOL, V262, P522
[7]   DNA-POLYMERASES DELTA AND EPSILON ARE REQUIRED FOR CHROMOSOMAL REPLICATION IN SACCHAROMYCES-CEREVISIAE [J].
BUDD, ME ;
CAMPBELL, JL .
MOLECULAR AND CELLULAR BIOLOGY, 1993, 13 (01) :496-505
[8]  
CAMPBELL JL, 1993, J BIOL CHEM, V268, P25261
[9]   Cell cycle control of replication initiation in eukaryotes [J].
Chevalier, S ;
Blow, JJ .
CURRENT OPINION IN CELL BIOLOGY, 1996, 8 (06) :815-821
[10]   P25(RUM1) ORDERS S-PHASE AND MITOSIS BY ACTING AS AN INHIBITOR OF THE P34(CDC2) MITOTIC KINASE [J].
CORREABORDES, J ;
NURSE, P .
CELL, 1995, 83 (06) :1001-1009