Licensing regulators Geminin and Cdt1 identify progenitor cells of the mouse CNS in a specific phase of the cell cycle

Spella, Magda; Britz, Olivier; Kotantaki, Panoraia; Lygerou, Zoi; Nishitani, Hideo; Ramsay, Robert G.; Flordellis, Christodoulos S.; Guillemot, François; Mantamadiotis, Theo; Taraviras, Stavros

doi:10.1016/j.neuroscience.2007.03.050

Cited by 42 publications

(57 citation statements)

References 59 publications

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“…In contrast, Geminin mRNA was localized throughout the ventricular and subventricular zones of the embryonic telencephalon (Fig. 7Ab) where proliferating cells are localized, consistent with previous studies (33).…”

Section: Figure 3 Idas Directly Binds To Geminin In Vitro and Prevensupporting

confidence: 91%

“…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%

“…For control sense probe generation, the pBluescript vector containing the mIdas open reading frame was linearized with KpnI and transcribed with T7 RNA polymerase. Geminin sense and antisense probes were as described previously (32,33). Sense probes were used as the control in all experiments.…”

Section: Methodsmentioning

confidence: 99%

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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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Section: Figure 3 Idas Directly Binds To Geminin In Vitro and Prevensupporting

confidence: 91%

Section: Methodsmentioning

confidence: 99%

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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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“…Gem was identified by functional screening in Xenopus early embryos on the basis of its ability to increase the size of the neural plate at the expense of nonneural cell types such as epidermis (14). Gem was likewise shown to be highly expressed in pluripotent Xenopus ectoderm and to be restricted to the neural precursor territory, while being downregulated in neural cells before primary neuron differentiation (14,20,28). Although the mechanistic basis of Geminin's ability to promote neural gene expression in Xenopus embryos has not been defined, our data suggest that this activity may be conserved between lower vertebrates and mammals.…”

Section: Discussionmentioning

confidence: 67%

Geminin promotes neural fate acquisition of embryonic stem cells by maintaining chromatin in an accessible and hyperacetylated state

Yellajoshyula

Patterson

Elitt

et al. 2011

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

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Formation of the complex vertebrate nervous system begins when pluripotent cells of the early embryo are directed to acquire a neural fate. Although cell intrinsic controls play an important role in this process, the molecular nature of this regulation is not well defined. Here we assessed the role for Geminin, a nuclear protein expressed in embryonic cells, during neural fate acquisition from mouse embryonic stem (ES) cells. Whereas Geminin knockdown does not affect the ability of ES cells to maintain or exit pluripotency, we found that it significantly impairs their ability to acquire a neural fate. Conversely, Geminin overexpression promotes neural gene expression, even in the presence of growth factor signaling that antagonizes neural transcriptional responses. These data demonstrate that Geminin's activity contributes to mammalian neural cell fate acquisition. We investigated the mechanistic basis of this phenomenon and found that Geminin maintains a hyperacetylated and open chromatin conformation at neural genes. Interestingly, recombinant Geminin protein also rapidly alters chromatin acetylation and accessibility even when Geminin is combined with nuclear extract and chromatin in vitro. Together, these data support a role for Geminin as a cell intrinsic regulator of neural fate acquisition that promotes expression of neural genes by regulating chromatin accessibility and histone acetylation.

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“…It is noteworthy to mention that the Geminin (green fluorescence) and Cdt1 (red fluorescence) proteins used to design the FUCCI system 23 were previously shown to be abundantly expressed by neural progenitors during early neurogenesis in mice 30 and in adult brain tissues 9,25 . Although the mKO2-hCdt1(30/120) construct was mainly used in the present study to follow the G1 phase with red fluorescence, the use of both constructs [mKO2-hCdt1(30/120) and mAG-hGem(1/110)] could be envisioned to allow the visualization of the major phases of the cell cycle (G 1 and S-G 2 /M) as well as the G 1 /S transition 23 .…”

mentioning

confidence: 99%

Cell Sorting of Neural Stem and Progenitor Cells from the Adult Mouse Subventricular Zone and Live-imaging of their Cell Cycle Dynamics

Daynac

Morizur

Kortulewski

et al. 2015

JoVE

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Neural stem cells (NSCs) in the subventricular zone of the lateral ventricles (SVZ) sustain olfactory neurogenesis throughout life in the mammalian brain. They successively generate transit amplifying cells (TACs) and neuroblasts that differentiate into neurons once they integrate the olfactory bulbs. Emerging fluorescent activated cell sorting (FACS) techniques have allowed the isolation of NSCs as well as their progeny and have started to shed light on gene regulatory networks in adult neurogenic niches. We report here a cell sorting technique that allows to follow and distinguish the cell cycle dynamics of the above-mentioned cell populations from the adult SVZ with a LeX/EGFR/CD24 triple staining. Isolated cells are then plated as adherent cells to explore in details their cell cycle progression by time-lapse video microscopy. To this end, we use transgenic Fluorescence Ubiquitination Cell Cycle Indicator (FUCCI) mice in which cells are red-fluorescent during G 1 phase due to a G 1 specific red-Cdt1 reporter. This method has recently revealed that proliferating NSCs progressively lengthen their G 1 phase during aging, leading to neurogenesis impairment. This method is easily transposable to other systems and could be of great interest for the study of the cell cycle dynamics of brain cells in the context of brain pathologies.

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Licensing regulators Geminin and Cdt1 identify progenitor cells of the mouse CNS in a specific phase of the cell cycle

Cited by 42 publications

References 59 publications

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

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

Geminin promotes neural fate acquisition of embryonic stem cells by maintaining chromatin in an accessible and hyperacetylated state

Cell Sorting of Neural Stem and Progenitor Cells from the Adult Mouse Subventricular Zone and Live-imaging of their Cell Cycle Dynamics

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