A globular complex formation by Nda1 and the other five members of the MCM protein family in fission yeast

Adachi, Yasuhisa; Usukura, Jiro; Yanagida, Mitsuhiro

doi:10.1046/j.1365-2443.1997.1350333.x

Cited by 124 publications

(93 citation statements)

References 61 publications

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“…Although helicase activity with all six MCM proteins has remained elusive, a subcomplex of Mcm4/6/7p is a helicase (6 -8). Similar to replicative helicases in other systems, Mcm2-7p appears to be ring shaped, consistent with electron microscope studies of Mcm2-7p and Mcm4/ 6/7p (9,10). An archaeal MCM protein also has helicase activity and is an oligomeric ring (11)(12)(13)(14).…”

supporting

confidence: 83%

Reconstitution of the Mcm2-7p Heterohexamer, Subunit Arrangement, and ATP Site Architecture

Davey

O'Donnell

2003

Journal of Biological Chemistry

166

228

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supporting

confidence: 83%

Reconstitution of the Mcm2-7p Heterohexamer, Subunit Arrangement, and ATP Site Architecture

Davey

O'Donnell

2003

Journal of Biological Chemistry

166

228

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“…A helicase activity was reported to be associated with a complex of human Mcm4p,Mcm6p,and Mcm7p (32). Six fission yeast MCM proteins form a globular complex in vivo (1). Our data suggest that the MCM complex might associate with the B2 element of ARS1, a potential DNA-unwinding element (41).…”

Section: Discussionmentioning

confidence: 76%

Assembly of a Complex Containing Cdc45p, Replication Protein A, and Mcm2p at Replication Origins Controlled by S-Phase Cyclin-Dependent Kinases and Cdc7p-Dbf4p Kinase

Zou

Stillman

2000

Molecular and Cellular Biology

296

265

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In Saccharomyces cerevisiae, replication origins are activated with characteristic timing during S phase. S-phase cyclin-dependent kinases (S-CDKs) and Cdc7p-Dbf4p kinase are required for origin activation throughout S phase. The activation of S-CDKs leads to association of Cdc45p with chromatin, raising the possibility that Cdc45p defines the assembly of a new complex at each origin. Here we show that both Cdc45p and replication protein A (RPA) bind to Mcm2p at the G 1 -S transition in an S-CDK-dependent manner. During S phase, Cdc45p associates with different replication origins at specific times. The origin associations of Cdc45p and RPA are mutually dependent, and both S-CDKs and Cdc7p-Dbf4p are required for efficient binding of Cdc45p to origins. These findings suggest that S-CDKs and Cdc7p-Dbf4p promote loading of Cdc45p and RPA onto a preformed prereplication complex at each origin with preprogrammed timing. The ARS1 association of Mcm2p, but not that of the origin recognition complex, is diminished by disruption of the B2 element of ARS1, a potential origin DNA-unwinding element. Cdc45p is required for recruiting DNA polymerase ␣ onto chromatin, and it associates with Mcm2p, RPA, and DNA polymerase only during S phase. These results suggest that the complex containing Cdc45p, RPA, and MCMs is involved in origin unwinding and assembly of replication forks at each origin.In eukaryotic cells, DNA replication initiates from multiple origins scattered along chromosomes, making duplication of large genomes within a single cell cycle possible. Every replication origin fires once and only once with a characteristic timing during the S phase. Although the cell-cycle control of DNA replication has been gradually unveiled, how DNA replication is initiated at chromosomal origins remains largely unknown (reviewed in references 16, 21, 46, and 57).Studies of Saccharomyces cerevisiae and other organisms have revealed that a prereplication complex (pre-RC) is assembled at each origin during late mitosis or early G 1 phase, the period when cyclin-dependent kinases (CDKs) are inactive (11,12,15,40,51,58,59). The origin recognition complex (ORC), Cdc6p, and the six minichromosome maintenance (MCM) proteins Mcm2p to -7p are present in the pre-RC (2,19,40,55,59,63). To start DNA synthesis, S-phase CDKs (S-CDKs) and Cdc7p-Dbf4p kinase need to be activated at the G 1 -S transition. Upon activation of these kinases, the pre-RC undergoes a transformation, and it is eventually converted to the post-RC and replication forks when initiation of DNA replication is completed (16,21,46,57). To understand how DNA replication is initiated at chromosomal origins, we sought to uncover how the complexes at origins change during initiation and how they interact with origins. Furthermore, we pursued the mechanisms by which the activating kinases are involved in this transition.In the well-characterized simian virus 40 (SV40) system, large T antigen is the origin recognition protein and also the helicase. Initiation of DNA replication from the...

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“…Electron microscope images of Mcm2-7 and GINS complexes individually have shown that both complexes form ring-shaped structures (28,44). These findings suggest that the two rings may stack on top of each other to form a common central channel that could surround ssDNA or dsDNA.…”

Section: Discussionmentioning

confidence: 98%

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

Moyer

Lewis

Botchan

2006

Proc. Natl. Acad. Sci. U.S.A.

627

605

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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 Cdc45͞Mcm2-7͞GINS complex is associated with an active ATPdependent 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.Drosophila ͉ ATPase

show abstract

A globular complex formation by Nda1 and the other five members of the MCM protein family in fission yeast

Cited by 124 publications

References 61 publications

Reconstitution of the Mcm2-7p Heterohexamer, Subunit Arrangement, and ATP Site Architecture

Reconstitution of the Mcm2-7p Heterohexamer, Subunit Arrangement, and ATP Site Architecture

Assembly of a Complex Containing Cdc45p, Replication Protein A, and Mcm2p at Replication Origins Controlled by S-Phase Cyclin-Dependent Kinases and Cdc7p-Dbf4p Kinase

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

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