Localization of MCM2-7, Cdc45, and GINS to the Site of DNA Unwinding during Eukaryotic DNA Replication

Pacek, Marcin; Tutter, Antonin V.; Kubota, Yumiko; Takisawa, Haruhiko; Walter, Johannes C.

doi:10.1016/j.molcel.2006.01.030

Cited by 326 publications

(317 citation statements)

References 34 publications

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“…Chromatin immunoprecipitation (ChIP) was performed as described (Pacek et al , 2006). Briefly, reaction samples were crosslinked with formaldehyde, sonicated to yield DNA fragments of roughly 100–500 bp, and immunoprecipitated with the indicated antibodies.…”

Section: Methodsmentioning

confidence: 99%

Recruitment and positioning determine the specific role of the XPF ‐ ERCC 1 endonuclease in interstrand crosslink repair

et al. 2017

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XPF‐ERCC1 is a structure‐specific endonuclease pivotal for several DNA repair pathways and, when mutated, can cause multiple diseases. Although the disease‐specific mutations are thought to affect different DNA repair pathways, the molecular basis for this is unknown. Here we examine the function of XPF‐ERCC1 in DNA interstrand crosslink (ICL) repair. We used Xenopus egg extracts to measure both ICL and nucleotide excision repair, and we identified mutations that are specifically defective in ICL repair. One of these separation‐of‐function mutations resides in the helicase‐like domain of XPF and disrupts binding to SLX4 and recruitment to the ICL. A small deletion in the same domain supports recruitment of XPF to the ICL, but inhibited the unhooking incisions most likely by disrupting a second, transient interaction with SLX4. Finally, mutation of residues in the nuclease domain did not affect localization of XPF‐ERCC1 to the ICL but did prevent incisions on the ICL substrate. Our data support a model in which the ICL repair‐specific function of XPF‐ERCC1 is dependent on recruitment, positioning and substrate recognition.

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Section: Methodsmentioning

confidence: 99%

Recruitment and positioning determine the specific role of the XPF ‐ ERCC 1 endonuclease in interstrand crosslink repair

et al. 2017

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“…However, the area spanned by six patches is rather broad to accommodate a single protein, and thus it is likely that multiple proteins bind to these patches. Thus, based on structural and genetic analyses, we propose that this region within GINS might mediate the assembly of proteins in an RPC (Calzada et al 2005;Gambus et al 2006;Pacek et al 2006).…”

Section: Conserved Surface Regions With Functional Significancementioning

confidence: 99%

“…In budding yeast, chromatin immunoprecipitation analysis identified the GINS complex as a part of the replisome that contains MCM2-7, Cdc45, Pols ␣ and , and other replication regulation factors (Calzada et al 2005). The vertebrate replisome at the paused fork also contained GINS, along with Pols ␣ and , MCM2-7, Cdc45, and MCM10 (Pacek et al 2006). Moreover, a proteomics study in budding yeast showed that the GINS complex allows the MCM helicase to stably associate with Cdc45 in replisome progression complexes (RPCs) during and after the initiation of replication (Gambus et al 2006;Kanemaki and Labib 2006).…”

mentioning

confidence: 99%

Crystal structure of the human GINS complex

Choi¹,

Lim²,

Kim³

et al. 2007

Genes Dev.

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The GINS complex mediates the assembly of the MCM2-7 (minichromosome maintenance) complex with proteins in a replisome progression complex. The eukaryotic GINS complex is composed of Sld5, Psf1, Psf2, and Psf3, which must be assembled for cell proliferation. We determined the crystal structure of the human GINS complex: GINS forms an elliptical shape with a small central channel. The structures of Sld5 and Psf2 resemble those of Psf1 and Psf3, respectively. In addition, the N-terminal and C-terminal domains of Sld5/ Psf1 are permuted in Psf2/Psf3, which suggests that the four proteins have evolved from a common ancestor. Using a structure-based mutational analysis, we identified the functionally critical surface regions of the GINS complex.Supplemental material is available at http://www.genesdev.org.

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“…After loading onto DNA, the MCM complex interacts with CDC45 and GINS to unwind DNA at the replication fork (Gambus et al, 2006;Moyer et al, 2006;Pacek et al, 2006;Chang et al, 2007). Each of the MCM2 to MCM7 proteins contains several highly conserved sequence features, including an ATP hydrolysis domain, an Arg finger motif, and a zinc-binding domain (Maiorano et al, 2006).…”

mentioning

confidence: 99%

Dynamic Localization of the DNA Replication Proteins MCM5 and MCM7 in Plants

et al. 2009

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Genome integrity in eukaryotes depends on licensing mechanisms that prevent loading of the minichromosome maintenance complex (MCM2-7) onto replicated DNA during S phase. Although the principle of licensing appears to be conserved across all eukaryotes, the mechanisms that control it vary, and it is not clear how licensing is regulated in plants. In this work, we demonstrate that subunits of the MCM2-7 complex are coordinately expressed during Arabidopsis (Arabidopsis thaliana) development and are abundant in proliferating and endocycling tissues, indicative of a role in DNA replication. We show that endogenous MCM5 and MCM7 proteins are localized in the nucleus during G1, S, and G2 phases of the cell cycle and are released into the cytoplasmic compartment during mitosis. We also show that MCM5 and MCM7 are topologically constrained on DNA and that the MCM complex is stable under high-salt conditions. Our results are consistent with a conserved replicative helicase function for the MCM complex in plants but not with the idea that plants resemble budding yeast by actively exporting the MCM complex from the nucleus to prevent unauthorized origin licensing and rereplication during S phase. Instead, our data show that, like other higher eukaryotes, the MCM complex in plants remains in the nucleus throughout most of the cell cycle and is only dispersed in mitotic cells.

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Localization of MCM2-7, Cdc45, and GINS to the Site of DNA Unwinding during Eukaryotic DNA Replication

Cited by 326 publications

References 34 publications

Recruitment and positioning determine the specific role of the XPF ‐ ERCC 1 endonuclease in interstrand crosslink repair

Recruitment and positioning determine the specific role of the XPF ‐ ERCC 1 endonuclease in interstrand crosslink repair

Crystal structure of the human GINS complex

Dynamic Localization of the DNA Replication Proteins MCM5 and MCM7 in Plants

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