Isolation of the Cdc45/Mcm2-7/GINS (CMG) complex, a candidate for the eukaryotic DNA replication fork helicase

被引：546

作者：

Moyer, Stephen E. ^{[1
]}

Lewis, Peter W. ^{[1
]}

Botchan, Michael R. ^{[1
]}

机构：

[1] Univ Calif Berkeley, Dept Mol & Cell Biol, Div Biochem & Mol Biol, Berkeley, CA 94720 USA

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2006年 / 103卷 / 27期

关键词：

Drosophila; ATPase;

D O I：

10.1073/pnas.0602400103

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：

The protein Cdc45 plays a critical but poorly understood role in the initiation and elongation stages of eukaryotic DNA replication. To study Cdc45's function in DNA replication, we purified Cdc45 protein from Drosophila embryo extracts by a combination of traditional and immunoaffinity chromatography steps and found that the protein exists in a stable, high-molecular-weight complex with the Mcm2-7 hexamer and the GINS tetramer. The purified Cdc45/Mcm2-7/GINS complex is associated with an active ATP-dependent DNA helicase function. RNA interference knock-down experiments targeting the GINS and Cdc45 components establish that the proteins are required for the S phase transition in Drosophila cells. The data suggest that this complex forms the core helicase machinery for eukaryotic DNA replication.

引用

页码：10236 / 10241

页数：6

共 46 条

[1] A globular complex formation by Nda1 and the other five members of the MCM protein family in fission yeast [J].

Adachi, Y ;

Usukura, J ;

Yanagida, M .

GENES TO CELLS, 1997, 2 (07) :467-479

[2] Differential assembly of Cdc45p and DNA polymerases at early and late origins of DNA replication [J].

Aparicio, OM ;

Stout, AM ;

Bell, SP .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (16) :9130-9135

[3] 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

[4] A double-hexamer archaeal minichromosome maintenance protein is an ATP-dependent DNA helicase [J].

Chong, JPJ ;

Hayashi, MK ;

Simon, MN ;

Xu, RM ;

Stillman, B .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (04) :1530-1535

[5] Drosophila Mcm10 interacts with members of the prereplication complex and is required for proper chromosome condensation [J].

Christensen, TW ;

Tye, BK .

MOLECULAR BIOLOGY OF THE CELL, 2003, 14 (06) :2206-2215

[6] Mcm2, but not RPA, is a component of the mammalian early G1-phase prereplication complex [J].

Dimitrova, DS ;

Todorov, IT ;

Melendy, T ;

Gilbert, DM .

JOURNAL OF CELL BIOLOGY, 1999, 146 (04) :709-722

[7] Eukaryotic MCM proteins: Beyond replication initiation [J].

Forsburg, SL .

MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS, 2004, 68 (01) :109-+

[8] GINS maintains association of Cdc45 with MCM in replisome progression complexes at eukaryotic DNA replication forks [J].

Gambus, A ;

Jones, RC ;

Sanchez-Diaz, A ;

Kanemaki, M ;

van Deursen, F ;

Edmondson, RD ;

Labib, K .

NATURE CELL BIOLOGY, 2006, 8 (04) :358-U41

[9] Polymorphism and double hexamer structure in the archaeal minichromosome maintenance (MCM) helicase from Methanobacterium thermoautotrophicum [J].

Gómez-Llorente, Y ;

Fletcher, RJ ;

Chen, XJS ;

Carazo, JM ;

Martín, CS .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2005, 280 (49) :40909-40915

[10] mcm5/cdc46-bob1 bypasses the requirement for the S phase activator Cdc7p [J].

Hardy, CFJ ;

Dryga, O ;

Seematter, S ;

Pahl, PMB ;

Sclafani, RA .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (07) :3151-3155

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