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Characterization of mouse embryonic stem cells without any G1 cyclins

ABSTRACT: We performed regular RNA-seq to understand G1 cyclins function in ES cells

ORGANISM(S): Mus musculus

PROVIDER: GSE90091 | GEO | 2017/02/07

SECONDARY ACCESSION(S): PRJNA354410

REPOSITORIES: GEO

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Cell cycle and G1 cyclins

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D-type cyclins determine the activity of DNA mismatch repair in the G1 and S phases of the cell cycle

Project description:The large majority of oxidative lesions occurring in the G1 phase of the cell cycle are repaired by base excision repair (BER) rather than mismatch repair (MMR) to avoid long resections that can lead to genomic instability and cell death. However, how cells choose BER over MMR is not yet understood. Here, we show that, during G1, D-type cyclins are recruited to sites of oxidative DNA damage in a PCNA- and p21-dependent manner. In turn, in a manner that is independent on CDK4/6 activity, D-type cyclins stabilize p21, which competes through its PCNA-interacting protein (PIP) box with MMR components for their binding to PCNA. This reduces MMR activity while allowing BER. At the G1/S transition, the AMBRA1-dependent degradation of D-type cyclins renders p21 susceptible to proteolysis via SKP2 and CDT2. These timely degradation events allow the proper binding of MMR proteins to PCNA enabling the repair of DNA replication errors. Thus, the expression of D-type cyclins limit MMR in G1, whereas their degradation is necessary for proper MMR function in S. Defects in these two regulatory mechanisms promote genome instability. The mass spectrometry raw files correspond to the affinity purifications of proximity labeled (turbo-ID) PCNA and CCND1 under various conditions.

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Cell cycle and G1 cyclins

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Budding yeast relies on qualitative G1 cyclin specificity to couple cell cycle progression with morphogenetic development

Project description:Two models have been put forward for cyclin-dependent kinase (Cdk) control of the cell cycle. In the qualitative model, cell cycle events are ordered by distinct substrate specificities associated with successive waves of G1, S and mitotic cyclins. Alternatively, the gradual quantitative rise of Cdk activity from G1 phase to mitosis could lead to ordered substrate phosphorylation at sequential thresholds. Here, we study the relative contributions of qualitative and quantitative Cdk control in the budding yeast S. cerevisiae. S-phase cyclins can be replaced by a single mitotic cyclin, albeit at the cost of reduced fitness. The single cyclin can in addition replace G1 cyclins to support ordered cell cycle progression, fulfilling key predictions of the quantitative model. However, single-cyclin cells fail to polarize or grow buds and thus cannot sustain proliferation. Our results suggest that budding yeast has become dependent on G1 cyclin specificity to couple cell cycle progression to essential morphogenetic events.

2021-08-23 | PXD022757 | Pride

G1 cyclins link proliferation, pluripotency and differentiation of embryonic stem cells.

Project description: Not available

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Genome-wide analysis of mitosis, early G1, late G1 and G2 in mouse pluripotent cells

Project description:We provide the genome-wide map of Esrrb binding in mitotic and asynchronous ES cells together with the Esrrb-induced transcriptomic changes in early G1, late G1 and G2.

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Cell cycle and G1 cyclins

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Alkalihalobacillus lehensis G1 strain:G1

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Ganaspis sp. G1 strain:G1

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Emergence of undifferentiated colonies from mouse embryonic stem cells undergoing differentiation by retinoic acid treatment.

Project description:Pluripotent mouse ES cells maintain their genomic stability via regular passage through the 2-cell-like state (2CLS) with a distinct pattern of gene expression. In a standard culture, only ~5% cells are in the 2CLS; thus, it is desirable to find conditions that facilitate cell transition through this state as a method for improving the quality of ES cells. Here we show that low concentration of retinoic acid (RA) in the presence of LIF increases the proportion of cells undergoing the 2CLS without causing differentiation. Colonies of undifferentiated cells with high contribution of 2CLS marker, Zscan4c, appear clearly by the day 7. These colonies consist of a mixture of three types of cells: (1) cells in the 2CLS, (2) regular pluripotent ES cells that express Pou5f1, Sox2, and Nanog, and (3) endodermal cells, which are located at the periphery of the colony. Cell tracking with LacZ-staining shows that descendants of 2CLS-cells represent the majority of cells in these colonies. Cells exposed to RA and then transferred to the regular medium retain pluripotency, judged by their high ALP staining, typical ES colony morphology, gene expression profile, and stable karyotype. These cells readily differentiate into derivatives of all 3 germ layers in embryoid bodies assay and yield a higher contribution to chimeras after the injection into blastocysts than regular ES cells. In conclusion, transient culture of ES in low concentration of RA could increase the number of ES passaging through 2CLS and thus improve ES pluripotent phenotype and performance in chimera experiments.

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