Human Geminin promotes pre-RC formation and DNA replication by stabilizing CDT1 in mitosis

Ballabeni, Andrea; Melixetian, Marina; Zamponi, Raffaella; Masiero, Laura; Marinoni, Federica; Helin, Kristian

doi:10.1038/sj.emboj.7600314

Cited by 130 publications

(175 citation statements)

References 49 publications

(85 reference statements)

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“…Cdt1 displays cell cycle-dependent chromatin binding that is important for proper licensing regulation. As observed previously 18,19 , association of Cdt1 with chromatin was strong in NIH 3T3 cells synchronized in the G1 phase of the cell cycle, a period of pre-RC formation (Fig. 4a).…”

Section: W T δ P E S T V E C T V E C T W T δ P E S T V E C T V E C T supporting

confidence: 55%

“…Cdt1 binds to chromatin in a cell cycle-dependent fashion with high occupancy in G1 phase and low in mitosis [18][19][20] . It is not fully understood how this cell cycle-specific binding is controlled, but cyclin-dependent kinase (CDK) activities are involved in inhibiting Cdt1 association with chromatin 18,20 . Restraining Cdt1 binding to chromatin might impair licensing activity; however, the functional significance of the cell cycle-dependent chromatin association of Cdt1 is not well understood.…”

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confidence: 99%

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A spontaneous Cdt1 mutation in 129 mouse strains reveals a regulatory domain restraining replication licensing

et al. 2013

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Cdt1 is required for loading the replicative DNA helicase MCM2/7, a process known as DNA replication licensing. Here we show that 129 mouse strains express a Cdt1 mutated allele with enhanced licensing activity. The mutation, named D 6 PEST, involves a six-amino acid deletion within a previously uncharacterized PEST-like domain. Cdt1 D 6 PEST and more extensive deletions exhibit increased re-replication and transformation activities that are independent of the Geminin and E3 ligase pathways. This PEST domain negatively regulates cell cycledependent chromatin recruitment of Cdt1 in G2/M phases of the cell cycle. Mass spectrometry analysis indicates that Cdt1 is phosphorylated at sites within the deleted PEST domain during mitosis. This study reveals a conserved new regulatory Cdt1 domain crucial for proper DNA licensing activity and suggests a mechanism by which the presence of Cdt1 in G2/M phases does not lead to premature origin licensing. These results also question the usage of 129 mouse strains for knockout analyses.

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Section: W T δ P E S T V E C T V E C T W T δ P E S T V E C T V E C T supporting

confidence: 55%

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confidence: 99%

A spontaneous Cdt1 mutation in 129 mouse strains reveals a regulatory domain restraining replication licensing

et al. 2013

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“…A second vector for in situ hybridization was generated by subcloning the mIdas open reading frame to pBluescript between SacI and KpnI sites. The following plasmids have been described elsewhere: GemininGFP (29), Geminin⌬90 -120GFP (30,31), Cdt1dhcRed (31), GemininHA (29), plasmids for mouse Geminin in situ probes (32,33).…”

Section: Methodsmentioning

confidence: 99%

Idas, a Novel Phylogenetically Conserved Geminin-related Protein, Binds to Geminin and Is Required for Cell Cycle Progression

Pefani

Dimaki

Spella

et al. 2011

Journal of Biological Chemistry

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Development and homeostasis of multicellular organisms relies on an intricate balance between cell proliferation and differentiation. Geminin regulates the cell cycle by directly binding and inhibiting the DNA replication licensing factor Cdt1. Geminin also interacts with transcriptional regulators of differentiation and chromatin remodelling factors, and its balanced interactions are implicated in proliferation-differentiation decisions during development. Here, we describe Idas (Idas being a cousin of the Gemini in Ancient Greek Mythology), a previously uncharacterised coiled-coil protein related to Geminin. We show that human Idas localizes to the nucleus, forms a complex with Geminin both in cells and in vitro through coiled-coil mediated interactions, and can change Geminin subcellular localization. Idas does not associate with Cdt1 and prevents Geminin from binding to Cdt1 in vitro. Idas depletion from cells affects cell cycle progression; cells accumulate in S phase and are unable to efficiently progress to mitosis. Idas protein levels decrease in anaphase, whereas its overexpression causes mitotic defects. During development, we show that Idas exhibits high level expression in the choroid plexus and the cortical hem of the mouse telencephalon. Our data highlight Idas as a novel Geminin binding partner, implicated in cell cycle progression, and a putative regulator of proliferation-differentiation decisions during development.Development and homeostasis of multicellular organisms depends on the generation of the appropriate number of cells through cell proliferation and the acquisition of specialized cell functions through cell differentiation. Geminin has been suggested to regulate proliferation-differentiation decisions through balanced interactions with the DNA replication licensing factor Cdt1 and factors regulating transcription during development (1).

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“…The degradation of geminin by anaphasepromoting complex (APC) at the metaphase/anaphase transition, the dephosphorylation and the activation of replication factors, when the mitosis is completed, permits the initiation of a new round of DNA replication basal levels of Cdt1 during S phase and its accumulation during mitosis. 39 In addition, geminin can function as a negative or positive regulator of pre-RC formation in human cells and protect Cdt1 from proteasome-mediated degradation by inhibiting its ubiquitination. 39 …”

Section: Geminin-cdt1: An Important Balancementioning

confidence: 99%

“…39 In addition, geminin can function as a negative or positive regulator of pre-RC formation in human cells and protect Cdt1 from proteasome-mediated degradation by inhibiting its ubiquitination. 39 …”

Section: Geminin-cdt1: An Important Balancementioning

confidence: 99%

Role of geminin: from normal control of DNA replication to cancer formation and progression?

Montanari

Macaluso

Cittadini³

et al. 2006

Cell Death Differ

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The normal cell proliferation involves the passage through a series of finely regulated phases that are known as cell cycle checkpoints. In order to carry out a correct replication, the eukaryotic cell undergoes to a series of events ensuring that each copy of duplicated chromosome exactly segregates in each of daughter cells. 1,2 A main critical control of the cell cycle is to ensure that the DNA replication takes place and that it is followed by mitosis, once for each cell cycle. Alterations in the expression of genes that regulate the cell cycle can lead to malignant transformation and tumor progression by perturbing cell proliferation and/ or genomic stability. 3,4 The normal progression through the cell cycle is a critical step for the survival of eukaryotic cells, in which the initiation of DNA replication is under the tight control of several factors that ensure the exact duplication of chromosomes in S phase and their subsequent segregation in phase M. The strict regulation of the cell cycle combined with the presence of a licensing phase, in which the chromatin becomes licensed to be replicated, leads to the existence of a single DNA replication start site for each round of replication. 5 In this complicated network of signals regulating the cell cycle and maintaining genome integrity, the geminin protein is considered one of the main players. However, if the geminin acts as oncosuppressor gene or as a proto-oncogene is still an open question.In this article, we will report the mechanism by which geminin controls that the huge quantity of eukaryotes DNA, distributed over multiples chromosomes, is replicated once and once only for each cell cycle. Moreover, we will open and discuss an important question: what role does geminin play in cancer formation and progression? Does geminin act as an oncosuppressor, as a proto-oncogene or exhibit both roles? Geminin in the LicensingThe replication of eukaryotic genome is a complex process strictly regulated by an intricate intracellular and extracellular signaling network, which controls different phenomena such as cell proliferation, differentiation and apoptosis. 6,7 All these processes have a common function: the regulation of the initial phase of DNA replication. During this event, specific proteins sequentially assemble onto the replication origin by forming specific replicative complexes (pre-RC), which allow for the chromatin to be competent (licensed) for the next DNA replication step of the cell cycle. 8 Geminin is a 25 kDa nuclear protein that functions by inhibiting DNA replication. 9 During specific phases of the cell cycle, geminin is able to bind to Cdt1 protein and inhibits pre-RC formation. 10 In eukaryotes, the origins of replication are bound by a complex of six proteins, evolutionally well conserved among different species. 11,12 These proteins constitute the origin recognition complex (ORC) (1-6), a complex of proteins essential for the initiation of DNA replication, which was first identified in Saccharomyces cerevisiae as protein that bound t...

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Human Geminin promotes pre-RC formation and DNA replication by stabilizing CDT1 in mitosis

Cited by 130 publications

References 49 publications

A spontaneous Cdt1 mutation in 129 mouse strains reveals a regulatory domain restraining replication licensing

A spontaneous Cdt1 mutation in 129 mouse strains reveals a regulatory domain restraining replication licensing

Idas, a Novel Phylogenetically Conserved Geminin-related Protein, Binds to Geminin and Is Required for Cell Cycle Progression

Role of geminin: from normal control of DNA replication to cancer formation and progression?

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