Multi-step Loading of Human Minichromosome Maintenance Proteins in Live Human Cells

Symeonidou, Ioanna Eleni; Kotsantis, Panagiotis; Roukos, Vassilis; Rapsomaniki, Maria Anna; Grecco, Hernán E.; Bastiaens, Philippe I. H.; Taraviras, Stavros; Lygerou, Zoi

doi:10.1074/jbc.m113.474825

Cited by 34 publications

(30 citation statements)

References 69 publications

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“…34 The mechanistic implications of elevated expression and potential strategies to intervene, hinge on a detailed knowledge of the process as it occurs in mammalian cells. Previous work in yeast, 35 Xenopus, 4 cancer cells, 36,37 and CHO cells, 38 all demonstrate stable immobilisation of MCM2 in the nucleus during G1 phase, however these analyses do not further define the binding properties over this crucial period or distinguish NM-bound MCM from chromatin-bound MCM.…”

Section: Discussionmentioning

confidence: 99%

Transient association of MCM complex proteins with the nuclear matrix during initiation of mammalian DNA replication

et al. 2015

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The minichromosome maintenance complex (MCM2-7) is the putative DNA helicase in eukaryotes, and essential for DNA replication. By applying serial extractions to mammalian cells synchronized by release from quiescence, we reveal dynamic changes to the sub-nuclear compartmentalization of MCM2 as cells pass through late G1 and early S phase, identifying a brief window when MCM2 becomes transiently attached to the nuclear-matrix. The data distinguish 3 states that correspond to loose association with chromatin prior to DNA replication, transient highly stable binding to the nuclear-matrix coincident with initiation, and a post-initiation phase when MCM2 remains tightly associated with chromatin but not the nuclear-matrix. The data suggests that functional MCM complex loading takes place at the nuclear-matrix.

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Section: Discussionmentioning

confidence: 99%

Transient association of MCM complex proteins with the nuclear matrix during initiation of mammalian DNA replication

et al. 2015

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“…35 Thus, it is possible that a continuous supply of nascent MCM2-7 might be necessary to maintain the licensed state during the long G1 phase in the somatic cell cycle. 36 Overall, the use of maternal MCM2-7 complexes only during the embryonic cell cycle, which are defective in nuclear transport activity, seems to have a specific role.…”

Section: Discussionmentioning

confidence: 99%

Evolutionary diversification ofMCM3genes inXenopus laevisandDanio rerio

et al. 2014

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Embryonic cell cycles of amphibians are rapid and lack zygotic transcription and checkpoint control. At the midblastula transition, zygotic transcription is initiated and cell divisions become asynchronous. Several cell cycle-related amphibian genes retain 2 distinct forms, maternal and zygotic, but little is known about the functional differences between these 2 forms of proteins. The minichromosome maintenance (MCM) 2-7 complex, consisting of 6 MCM proteins, plays a central role in the regulation of eukaryotic DNA replication. Almost all eukaryotes retain just a single MCM gene for each subunit. Here we report that Xenopus and zebrafish have 2 copies of MCM3 genes, one of which shows a maternal and the other a zygotic expression pattern. Phylogenetic analysis shows that the Xenopus and zebrafish zygotic MCM3 genes are more similar to their mammalian MCM3 ortholog, suggesting that maternal MCM3 was lost during evolution in most vertebrate lineages. Maternal MCM3 proteins in these 2 species are functionally different from zygotic MCM3 proteins because zygotic, but not maternal, MCM3 possesses an active nuclear localization signal in its C-terminal region, such as mammalian MCM3 orthologs do. mRNA injection experiments in zebrafish embryos show that overexpression of maternal MCM3 impairs proliferation and causes developmental defects, whereas zygotic MCM3 has a much weaker effect. This difference is brought about by the difference in their C-terminal regions, which contain putative nuclear localization signals; swapping the C-terminal region between maternal and zygotic genes diminishes the developmental defects. This study suggests that evolutionary diversification has occurred in MCM3 genes, leading to distinct functions, possibly as an adaption to the rapid DNA replication required for early development of Xenopus and zebrafish.

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“…The binding of Geminin to Cdt1 inhibits the loading of the mini-chromosome maintenance complex (MCM) onto chromatin and pre-replication complex (preRC) formation (Tada et al, 2001;Wohlschlegel et al, 2000;reviewed in Lygerou & Nurse, 2000;Symeonidou et al, 2013). Besides its role in proliferation, Geminin also has a role in cell differentiation (Seo & Kroll, 2006;Champeris Tsaniras et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

The structure of the GemC1 coiled coil and its interaction with the Geminin family of coiled-coil proteins

Caillat

Fish

Pefani

et al. 2015

Acta Crystallogr D Biol Crystallogr

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GemC1, together with Idas and Geminin, an important regulator of DNA-replication licensing and differentiation decisions, constitute a superfamily sharing a homologous central coiled-coil domain. To better understand this family of proteins, the crystal structure of a GemC1 coiled-coil domain variant engineered for better solubility was determined to 2.2 Å resolution. GemC1 shows a less typical coiled coil compared with the Geminin homodimer and the Geminin–Idas heterodimer structures. It is also shown that both in vitro and in cells GemC1 interacts with Geminin through its coiled-coil domain, forming a heterodimer that is more stable that the GemC1 homodimer. Comparative analysis of the thermal stability of all of the possible superfamily complexes, using circular dichroism to follow the unfolding of the entire helix of the coiled coil, or intrinsic tryptophan fluorescence of a unique conserved N-terminal tryptophan, shows that the unfolding of the coiled coil is likely to take place from the C-terminus towards the N-terminus. It is also shown that homodimers show a single-state unfolding, while heterodimers show a two-state unfolding, suggesting that the dimer first falls apart and the helices then unfold according to the stability of each protein. The findings argue that Geminin-family members form homodimers and heterodimers between them, and this ability is likely to be important for modulating their function in cycling and differentiating cells.

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Multi-step Loading of Human Minichromosome Maintenance Proteins in Live Human Cells

Cited by 34 publications

References 69 publications

Transient association of MCM complex proteins with the nuclear matrix during initiation of mammalian DNA replication

Transient association of MCM complex proteins with the nuclear matrix during initiation of mammalian DNA replication

Evolutionary diversification ofMCM3genes inXenopus laevisandDanio rerio

The structure of the GemC1 coiled coil and its interaction with the Geminin family of coiled-coil proteins

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