Phosphoproteomics identifies a bimodal EPHA2 receptor switch that promotes embryonic stem cell differentiation

Rosalia Fernandez-Alonso; Francisco Bustos; Manon Budzyk; Pankaj Kumar; Andreas O. Helbig; Jens Hukelmann; Angus I. Lamond; Fredrik Lanner; Houjiang Zhou; Evangelia Petsalaki; Greg M. Findlay

doi:10.1038/s41467-020-15173-4

Phosphoproteomics identifies a bimodal EPHA2 receptor switch that promotes embryonic stem cell differentiation

Rosalia Fernandez-Alonso, Francisco Bustos, Manon Budzyk, Pankaj Kumar, Andreas O. Helbig, Jens Hukelmann, Angus I. Lamond, Fredrik Lanner, Houjiang Zhou, Evangelia Petsalaki, Greg M. Findlay (Lead / Corresponding author)

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Abstract

Embryonic Stem Cell (ESC) differentiation requires complex cell signalling network dynamics, although the key molecular events remain poorly understood. Here, we use phosphoproteomics to identify an FGF4-mediated phosphorylation switch centred upon the key Ephrin receptor EPHA2 in differentiating ESCs. We show that EPHA2 maintains pluripotency and restrains commitment by antagonising ERK1/2 signalling. Upon ESC differentiation, FGF4 utilises a bimodal strategy to disable EPHA2, which is accompanied by transcriptional induction of EFN ligands. Mechanistically, FGF4-ERK1/2-RSK signalling inhibits EPHA2 via Ser/Thr phosphorylation, whilst FGF4-ERK1/2 disrupts a core pluripotency transcriptional circuit required for Epha2 gene expression. This system also operates in mouse and human embryos, where EPHA receptors are enriched in pluripotent cells whilst surrounding lineage-specified trophectoderm expresses EFNA ligands. Our data provide insight into function and regulation of EPH-EFN signalling in ESCs, and suggest that segregated EPH-EFN expression coordinates cell fate with compartmentalisation during early embryonic development.

Original language	English
Article number	1357
Number of pages	13
Journal	Nature Communications
Volume	11
DOIs	https://doi.org/10.1038/s41467-020-15173-4
Publication status	Published - 13 Mar 2020

Keywords

Cell signalling
Embryonic stem cells
Phosphorylation
Proteomics

ASJC Scopus subject areas

General Physics and Astronomy
General Chemistry
General Biochemistry,Genetics and Molecular Biology

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10.1038/s41467-020-15173-4Licence: CC BY

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Cite this

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title = "Phosphoproteomics identifies a bimodal EPHA2 receptor switch that promotes embryonic stem cell differentiation",

abstract = "Embryonic Stem Cell (ESC) differentiation requires complex cell signalling network dynamics, although the key molecular events remain poorly understood. Here, we use phosphoproteomics to identify an FGF4-mediated phosphorylation switch centred upon the key Ephrin receptor EPHA2 in differentiating ESCs. We show that EPHA2 maintains pluripotency and restrains commitment by antagonising ERK1/2 signalling. Upon ESC differentiation, FGF4 utilises a bimodal strategy to disable EPHA2, which is accompanied by transcriptional induction of EFN ligands. Mechanistically, FGF4-ERK1/2-RSK signalling inhibits EPHA2 via Ser/Thr phosphorylation, whilst FGF4-ERK1/2 disrupts a core pluripotency transcriptional circuit required for Epha2 gene expression. This system also operates in mouse and human embryos, where EPHA receptors are enriched in pluripotent cells whilst surrounding lineage-specified trophectoderm expresses EFNA ligands. Our data provide insight into function and regulation of EPH-EFN signalling in ESCs, and suggest that segregated EPH-EFN expression coordinates cell fate with compartmentalisation during early embryonic development.",

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author = "Rosalia Fernandez-Alonso and Francisco Bustos and Manon Budzyk and Pankaj Kumar and Helbig, {Andreas O.} and Jens Hukelmann and Lamond, {Angus I.} and Fredrik Lanner and Houjiang Zhou and Evangelia Petsalaki and Findlay, {Greg M.}",

note = "Funding: MRC New Investigator Research Grant (MR/N000609/1); Wellcome Trust/Royal Society Sir Henry Dale Fellowship (211209/Z/18/Z)",

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doi = "10.1038/s41467-020-15173-4",

language = "English",

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T1 - Phosphoproteomics identifies a bimodal EPHA2 receptor switch that promotes embryonic stem cell differentiation

AU - Fernandez-Alonso, Rosalia

AU - Bustos, Francisco

AU - Budzyk, Manon

AU - Kumar, Pankaj

AU - Helbig, Andreas O.

AU - Hukelmann, Jens

AU - Lamond, Angus I.

AU - Lanner, Fredrik

AU - Zhou, Houjiang

AU - Petsalaki, Evangelia

AU - Findlay, Greg M.

N1 - Funding: MRC New Investigator Research Grant (MR/N000609/1); Wellcome Trust/Royal Society Sir Henry Dale Fellowship (211209/Z/18/Z)

PY - 2020/3/13

Y1 - 2020/3/13

N2 - Embryonic Stem Cell (ESC) differentiation requires complex cell signalling network dynamics, although the key molecular events remain poorly understood. Here, we use phosphoproteomics to identify an FGF4-mediated phosphorylation switch centred upon the key Ephrin receptor EPHA2 in differentiating ESCs. We show that EPHA2 maintains pluripotency and restrains commitment by antagonising ERK1/2 signalling. Upon ESC differentiation, FGF4 utilises a bimodal strategy to disable EPHA2, which is accompanied by transcriptional induction of EFN ligands. Mechanistically, FGF4-ERK1/2-RSK signalling inhibits EPHA2 via Ser/Thr phosphorylation, whilst FGF4-ERK1/2 disrupts a core pluripotency transcriptional circuit required for Epha2 gene expression. This system also operates in mouse and human embryos, where EPHA receptors are enriched in pluripotent cells whilst surrounding lineage-specified trophectoderm expresses EFNA ligands. Our data provide insight into function and regulation of EPH-EFN signalling in ESCs, and suggest that segregated EPH-EFN expression coordinates cell fate with compartmentalisation during early embryonic development.

AB - Embryonic Stem Cell (ESC) differentiation requires complex cell signalling network dynamics, although the key molecular events remain poorly understood. Here, we use phosphoproteomics to identify an FGF4-mediated phosphorylation switch centred upon the key Ephrin receptor EPHA2 in differentiating ESCs. We show that EPHA2 maintains pluripotency and restrains commitment by antagonising ERK1/2 signalling. Upon ESC differentiation, FGF4 utilises a bimodal strategy to disable EPHA2, which is accompanied by transcriptional induction of EFN ligands. Mechanistically, FGF4-ERK1/2-RSK signalling inhibits EPHA2 via Ser/Thr phosphorylation, whilst FGF4-ERK1/2 disrupts a core pluripotency transcriptional circuit required for Epha2 gene expression. This system also operates in mouse and human embryos, where EPHA receptors are enriched in pluripotent cells whilst surrounding lineage-specified trophectoderm expresses EFNA ligands. Our data provide insight into function and regulation of EPH-EFN signalling in ESCs, and suggest that segregated EPH-EFN expression coordinates cell fate with compartmentalisation during early embryonic development.

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Phosphoproteomics identifies a bimodal EPHA2 receptor switch that promotes embryonic stem cell differentiation

Abstract

Keywords

ASJC Scopus subject areas

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Functions and Applications of a Novel Pluripotency Signalling Pathway (Sir Henry Dale Fellowship)

Elucidating Novel Pluripotency Signalling Networks (New Investigator Award)

Findlay, Greg

Cite this

Phosphoproteomics identifies a bimodal EPHA2 receptor switch that promotes embryonic stem cell differentiation

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Projects

Functions and Applications of a Novel Pluripotency Signalling Pathway (Sir Henry Dale Fellowship)

Elucidating Novel Pluripotency Signalling Networks (New Investigator Award)

Profiles

Findlay, Greg

Cite this