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Mus musculus

ABSTRACT: RISC-mediated control of selected chromatin regulators stabilizes ground state pluripotency of mouse embryonic stem cells (TRAP)

PROVIDER: PRJNA316606 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR3306081.fastq.gz	Fastqsanger.gz
	SRR3306082.fastq.gz	Fastqsanger.gz
	SRR3306083.fastq.gz	Fastqsanger.gz
	SRR3306084.fastq.gz	Fastqsanger.gz
	SRR3306085.fastq.gz	Fastqsanger.gz

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RISC-mediated control of selected chromatin regulators stabilizes ground state pluripotency of mouse embryonic stem cells

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PRAMEL7 and CUL2 decrease NuRD stability to establish ground-state pluripotency

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Spic a specific marker of ground state pluripotency that regulates one carbon metabolism

Project description:Understanding mechanisms of epigenetic regulation in embryonic stem cells (ESCs) is of fundamental importance for stem cell and developmental biology. Here we identify Spic, a member of the ETS family of transcription factors, as a specific marker of ground state pluripotency. We show that Spic is rapidly induced in ESCs cultured with GSK3-, MEK-inhibitors and LIF (2iL), and in response to MEK/ERK inhibition. ChIP-seq analysis demonstrated that Spic binds to enhancer elements that are associated with pluripotency genes. Interaction proteomics and genomic profiling confirmed that SPIC interacts with NANOG and stabilizes its binding to chromatin in 2iL-ESCs. Additional gain of function and loss of function experiments revealed that Spic controls genes involved in one carbon (1C) metabolism, Bhmt, Bhmt2, and Dmgdh, and the flux of SAM-to-SAH in 2iL-ESCs. By maintaining low levels of SAM, Spic controls the level of H3K4me3 and H3R17me2 histone methylation in ground state ESCs. Our data highlight the role of uncharacterized axillary transcription factors that link cellular metabolism to epigenetic regulation in ground state pluripotency.

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RISC-mediated control of selected chromatin regulators stabilizes ground state pluripotency of mouse embryonic stem cells (TRAP)

Project description:We performed an integrated analysis of RNA and proteins at the transition between naÃ¯ve ES cells and cells primed to differentiate. During this transition, mRNAs coding for chromatin regulators were specifically released from translational inhibition mediated by RNA-Induced Silencing Complex (RISC). This suggests that, prior to differentiation, the propensity of ES cells to change their epigenetic status is hampered by RNA interference. The expression of these chromatin regulators was reinstated following acute inactivation of RISC, and it correlated with loss of stemness markers and activation of early cell differentiation markers in treated ES cells. We evaluated ribosome occupancy of mRNAs in embryonic stem cells and Epiblast-Like Aggregates by means of polysome profiling with a modified Translating Ribosome Affinity Purification protocol (TRAP).

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(RNA-seq) Spic regulates one-carbon metabolism and histone methylation in ground-state pluripotency

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