Phosphorylation of NANOG by casein kinase I regulates embryonic stem cell self-renewal

Nicholas P. Mullin; Joby Varghese; Douglas Colby; Julia M. Richardson; Greg M. Findlay; Ian Chambers

doi:10.1002/1873-3468.13969

Phosphorylation of NANOG by casein kinase I regulates embryonic stem cell self-renewal

Nicholas P. Mullin, Joby Varghese, Douglas Colby, Julia M. Richardson, Greg M. Findlay, Ian Chambers (Lead / Corresponding author)

MRC PPU

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Abstract

The self-renewal efficiency of mouse embryonic stem cells (ESCs) is determined by the concentration of the transcription factor NANOG. While NANOG binds thousands of sites in chromatin, the regulatory systems that control DNA binding are poorly characterised. Here, we show that NANOG is phosphorylated by casein kinase I, and identify target residues. Phosphomimetic substitutions at phosphorylation sites within the homeodomain (S130 and S131) have site-specific functional effects. Phosphomimetic substitution of S130 abolishes DNA binding by NANOG and eliminates LIF-independent self-renewal. In contrast, phosphomimetic substitution of S131 enhances LIF-independent self-renewal, without influencing DNA binding. Modelling the DNA–homeodomain complex explains the disparate effects of these phosphomimetic substitutions. These results indicate how phosphorylation may influence NANOG homeodomain interactions that underpin ESC self-renewal.

Original language	English
Pages (from-to)	14-25
Number of pages	12
Journal	FEBS Letters
Volume	595
Issue number	1
Early online date	26 Oct 2020
DOIs	https://doi.org/10.1002/1873-3468.13969
Publication status	Published - Jan 2021

Keywords

DNA binding
NANOG
casein kinase I
phosphorylation
self-renewal

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1002/1873-3468.13969

Final Published VersionFinal published version, 1.55 MBLicence: CC BY

Cite this

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title = "Phosphorylation of NANOG by casein kinase I regulates embryonic stem cell self-renewal",

abstract = "The self-renewal efficiency of mouse embryonic stem cells (ESCs) is determined by the concentration of the transcription factor NANOG. While NANOG binds thousands of sites in chromatin, the regulatory systems that control DNA binding are poorly characterised. Here, we show that NANOG is phosphorylated by casein kinase I, and identify target residues. Phosphomimetic substitutions at phosphorylation sites within the homeodomain (S130 and S131) have site-specific functional effects. Phosphomimetic substitution of S130 abolishes DNA binding by NANOG and eliminates LIF-independent self-renewal. In contrast, phosphomimetic substitution of S131 enhances LIF-independent self-renewal, without influencing DNA binding. Modelling the DNA–homeodomain complex explains the disparate effects of these phosphomimetic substitutions. These results indicate how phosphorylation may influence NANOG homeodomain interactions that underpin ESC self-renewal.",

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note = "Funding Information: Research in IC{\textquoteright}s lab is supported by the Medical Research Council and the Biotechnology and Biological Sciences Research Council of the UK. Work in GF{\textquoteright}s laboratory is supported by a Medical Research Council New Investigator Research Grant and a Wellcome Trust/Royal Society Sir Henry Dale Fellowship. JV is supported by the Medical Research Council Protein Phosphorylation and Ubiquitylation Unit core grant. Publisher Copyright: {\textcopyright} 2020 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies",

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AU - Mullin, Nicholas P.

AU - Varghese, Joby

AU - Colby, Douglas

AU - Richardson, Julia M.

AU - Findlay, Greg M.

AU - Chambers, Ian

N1 - Funding Information: Research in IC’s lab is supported by the Medical Research Council and the Biotechnology and Biological Sciences Research Council of the UK. Work in GF’s laboratory is supported by a Medical Research Council New Investigator Research Grant and a Wellcome Trust/Royal Society Sir Henry Dale Fellowship. JV is supported by the Medical Research Council Protein Phosphorylation and Ubiquitylation Unit core grant. Publisher Copyright: © 2020 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies

PY - 2021/1

Y1 - 2021/1

N2 - The self-renewal efficiency of mouse embryonic stem cells (ESCs) is determined by the concentration of the transcription factor NANOG. While NANOG binds thousands of sites in chromatin, the regulatory systems that control DNA binding are poorly characterised. Here, we show that NANOG is phosphorylated by casein kinase I, and identify target residues. Phosphomimetic substitutions at phosphorylation sites within the homeodomain (S130 and S131) have site-specific functional effects. Phosphomimetic substitution of S130 abolishes DNA binding by NANOG and eliminates LIF-independent self-renewal. In contrast, phosphomimetic substitution of S131 enhances LIF-independent self-renewal, without influencing DNA binding. Modelling the DNA–homeodomain complex explains the disparate effects of these phosphomimetic substitutions. These results indicate how phosphorylation may influence NANOG homeodomain interactions that underpin ESC self-renewal.

AB - The self-renewal efficiency of mouse embryonic stem cells (ESCs) is determined by the concentration of the transcription factor NANOG. While NANOG binds thousands of sites in chromatin, the regulatory systems that control DNA binding are poorly characterised. Here, we show that NANOG is phosphorylated by casein kinase I, and identify target residues. Phosphomimetic substitutions at phosphorylation sites within the homeodomain (S130 and S131) have site-specific functional effects. Phosphomimetic substitution of S130 abolishes DNA binding by NANOG and eliminates LIF-independent self-renewal. In contrast, phosphomimetic substitution of S131 enhances LIF-independent self-renewal, without influencing DNA binding. Modelling the DNA–homeodomain complex explains the disparate effects of these phosphomimetic substitutions. These results indicate how phosphorylation may influence NANOG homeodomain interactions that underpin ESC self-renewal.

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Phosphorylation of NANOG by casein kinase I regulates embryonic stem cell self-renewal

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)

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Phosphorylation of NANOG by casein kinase I regulates embryonic stem cell self-renewal

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)

Cite this