BMP Sustains Embryonic Stem Cell Self-Renewal through Distinct Functions of Different Krüppel-like Factors

Masato Morikawa; Daizo Koinuma; Anna Mizutani; Natsumi Kawasaki; Katarina Holmborn; Anders Sundqvist; Shuichi Tsutsumi; Tetsuro Watabe; Hiroyuki Aburatani; Carl-Henrik Heldin; Kohei Miyazono

doi:10.1016/j.stemcr.2015.12.004

BMP Sustains Embryonic Stem Cell Self-Renewal through Distinct Functions of Different Krüppel-like Factors

Stem Cell Reports. 2016 Jan 12;6(1):64-73. doi: 10.1016/j.stemcr.2015.12.004.

Affiliations

¹ Ludwig Institute for Cancer Research, Science for Life Laboratory, Uppsala University, 751 24 Uppsala, Sweden.
² Department of Molecular Pathology, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan.
³ Zebrafish Technology Platform, Science for Life Laboratory, Uppsala University, 752 36 Uppsala, Sweden.
⁴ Genome Science Division, Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo 153-8904, Japan.
⁵ Department of Molecular Pathology, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan; JST, PRESTO, Kawaguchi, Saitama 332-0012, Japan.
⁶ Ludwig Institute for Cancer Research, Science for Life Laboratory, Uppsala University, 751 24 Uppsala, Sweden. Electronic address: c-h.heldin@licr.uu.se.
⁷ Ludwig Institute for Cancer Research, Science for Life Laboratory, Uppsala University, 751 24 Uppsala, Sweden; Department of Molecular Pathology, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan. Electronic address: miyazono@m.u-tokyo.ac.jp.

Abstract

Bone morphogenetic protein (BMP) signaling exerts paradoxical roles in pluripotent stem cells (PSCs); it sustains self-renewal of mouse embryonic stem cells (ESCs), while it induces differentiation in other PSCs, including human ESCs. Here, we revisit the roles of BMP-4 using mouse ESCs (mESCs) in naive and primed states. SMAD1 and SMAD5, which transduce BMP signals, recognize enhancer regions together with KLF4 and KLF5 in naive mESCs. KLF4 physically interacts with SMAD1 and suppresses its activity. Consistently, a subpopulation of cells with active BMP-SMAD can be ablated without disturbing the naive state of the culture. Moreover, Smad1/5 double-knockout mESCs stay in the naive state, indicating that the BMP-SMAD pathway is dispensable for it. In contrast, the MEK5-ERK5 pathway mediates BMP-4-induced self-renewal of mESCs by inducing Klf2, a critical factor for the ground state pluripotency. Our study illustrates that BMP exerts its self-renewing effect through distinct functions of different Krüppel-like factors.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Blotting, Western
Bone Morphogenetic Protein 4 / genetics
Bone Morphogenetic Protein 4 / metabolism*
Cell Differentiation / genetics
Cell Differentiation / physiology
Cell Self Renewal / genetics
Cell Self Renewal / physiology*
Gene Expression Regulation, Developmental
HEK293 Cells
Hep G2 Cells
Humans
Kruppel-Like Factor 4
Kruppel-Like Transcription Factors / genetics
Kruppel-Like Transcription Factors / metabolism*
MAP Kinase Signaling System / genetics
MAP Kinase Signaling System / physiology
Mice
Mice, Knockout
Mouse Embryonic Stem Cells / cytology
Mouse Embryonic Stem Cells / metabolism*
Pluripotent Stem Cells / cytology
Pluripotent Stem Cells / metabolism
Protein Binding
Reverse Transcriptase Polymerase Chain Reaction
Smad1 Protein / genetics
Smad1 Protein / metabolism
Smad5 Protein / genetics
Smad5 Protein / metabolism
Zebrafish / embryology
Zebrafish / genetics
Zebrafish / metabolism

Substances

Bone Morphogenetic Protein 4
KLF4 protein, human
Klf2 protein, mouse
Klf4 protein, mouse
Klf5 protein, mouse
Kruppel-Like Factor 4
Kruppel-Like Transcription Factors
Smad1 Protein
Smad5 Protein