Negative feedback via RSK modulates Erk‐dependent progression from naïve pluripotency

• Published in: EMBO reports Vol. 19; no. 8
• Main Authors: Nett, Isabelle RE, Mulas, Carla, Gatto, Laurent, Lilley, Kathryn S, Smith, Austin
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.08.2018; Springer Nature B.V; John Wiley and Sons Inc
• Subjects: Activation; Animals; Cell differentiation; Cell Differentiation - drug effects; Cell Lineage - drug effects; Combinatorial analysis; CRISPR; Differentiation (biology); EMBO31; EMBO37; EMBO39; embryonic stem cells; Embryonic Stem Cells - cytology; Embryonic Stem Cells - drug effects; Embryonic Stem Cells - metabolism; Extracellular signal-regulated kinase; Extracellular Signal-Regulated MAP Kinases - metabolism; Feedback; Feedback, Physiological - drug effects; Genotypes; Humans; Kinases; MAP kinase; mitogen‐activated protein kinase; Mutation; Mutation - genetics; Negative feedback; Organic chemistry; Phosphoproteins - metabolism; Phosphorylation; Phosphorylation - drug effects; Pluripotency; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - drug effects; Pluripotent Stem Cells - metabolism; Protein kinase; Proteins; Proteome - metabolism; Ribosomal protein S6 kinase; Ribosomal Protein S6 Kinases, 90-kDa - antagonists & inhibitors; Ribosomal Protein S6 Kinases, 90-kDa - genetics; Ribosomal Protein S6 Kinases, 90-kDa - metabolism; RSK; Signal transduction; Signaling; signalling feedback; Small Molecule Libraries - pharmacology; pluripotency; RSK; signalling feedback; mitogen‐activated protein kinase; embryonic stem cells
• ISSN: 1469-221X; 1469-3178; 1469-3178
• DOI: 10.15252/embr.201745642

Mitogen‐activated protein kinase (MAPK)/extracellular signal‐regulated kinase (ERK) signalling is implicated in initiation of embryonic stem (ES) cell differentiation. The pathway is subject to complex feedback regulation. Here, we examined the ERK‐responsive phosphoproteome in ES cells and identified the negative regulator RSK1 as a prominent target. We used CRISPR/Cas9 to create combinatorial mutations in RSK family genes. Genotypes that included homozygous null mutations in Rps6ka1, encoding RSK1, resulted in elevated ERK phosphorylation. These RSK‐depleted ES cells exhibit altered kinetics of transition into differentiation, with accelerated downregulation of naïve pluripotency factors, precocious expression of transitional epiblast markers and early onset of lineage specification. We further show that chemical inhibition of RSK increases ERK phosphorylation and expedites ES cell transition without compromising multilineage potential. These findings demonstrate that the ERK activation profile influences the dynamics of pluripotency progression and highlight the role of signalling feedback in temporal control of cell state transitions. Synopsis MAPK/ERK signalling promotes ES cell differentiation. Negative feedback on ERK1/2 signalling influences the rate at which ES cells exit from naïve pluripotency. Interference with feedback regulator RSK increases the speed and homogeneity of transition to differentiation. Negative regulator RSK1 is a major target of ERK signalling in ES cells. RSK feedback restricts ERK activation and constrains exit from naïve pluripotency. Genetic depletion of RSKs accelerates ES cell differentiation. Chemical inhibition of RSKs increases the rate and uniformity of ES cell transition. Graphical Abstract MAPK/ERK signalling promotes ES cell differentiation. Negative feedback on ERK1/2 signalling influences the rate at which ES cells exit from naïve pluripotency. Interference with feedback regulator RSK increases the speed and homogeneity of transition to differentiation.