Nanog binds to Smad1 and blocks bone morphogenetic protein-induced differentiation of embryonic stem cells

Atsushi Suzuki; Ángel Raya; Yasuhiko Kawakami; Masanobu Morita; Takaaki Matsui; Kinichi Nakashima; Fred H Gage; Concepción Rodríguez-Esteban; Juan Carlos Izpisúa Belmonte

doi:10.1073/pnas.0506945103

Nanog binds to Smad1 and blocks bone morphogenetic protein-induced differentiation of embryonic stem cells

Proc Natl Acad Sci U S A. 2006 Jul 5;103(27):10294-10299. doi: 10.1073/pnas.0506945103. Epub 2006 Jun 26.

Authors

Atsushi Suzuki^{1

2}, Ángel Raya^{1

2

3}, Yasuhiko Kawakami¹, Masanobu Morita^{1

2}, Takaaki Matsui¹, Kinichi Nakashima^{4

5}, Fred H Gage^{2

4}, Concepción Rodríguez-Esteban¹, Juan Carlos Izpisúa Belmonte^{6

2

3}

Affiliations

¹ *Gene Expression Laboratory.
² Stem Cell Research Center, and.
³ Institució Catalana de Recerca i Estudis Avançats and Center of Regenerative Medicine, Dr. Aiguader 80, 08003 Barcelona, Spain; and.
⁴ Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037.
⁵ **Laboratory of Molecular Neuroscience, Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma 630-0101, Japan.
⁶ *Gene Expression Laboratory, [email protected].

Abstract

ES cells represent a valuable model for investigating early embryo development and hold promise for future regenerative medicine strategies. The self-renewal of pluripotent mouse ES cells has been shown to require extrinsic stimulation by the bone morphogenetic protein (BMP) and leukemia inhibitory factor signaling pathways and the expression of the transcription factors Oct4 and Nanog. However, the network of interactions among extrinsic and intrinsic determinants of ES cell pluripotency is currently poorly understood. Here, we show that Nanog expression is up-regulated in mouse ES cells by the binding of T (Brachyury) and STAT3 to an enhancer element in the mouse Nanog gene. We further show that Nanog blocks BMP-induced mesoderm differentiation of ES cells by physically interacting with Smad1 and interfering with the recruitment of coactivators to the active Smad transcriptional complexes. Taken together, our findings illustrate the existence of ES cell-specific regulatory networks that underlie the maintenance of ES cell pluripotency and provide mechanistic insights into the role of Nanog in this process.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Base Sequence
Binding Sites
Bone Morphogenetic Proteins / pharmacology*
Cell Differentiation / drug effects*
Cells, Cultured
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism*
Down-Regulation
Embryo, Mammalian / cytology*
Enhancer Elements, Genetic
Fetal Proteins / genetics
Fetal Proteins / metabolism
Homeodomain Proteins / genetics
Homeodomain Proteins / metabolism*
Mesoderm / cytology
Mesoderm / drug effects
Mesoderm / metabolism
Mice
Molecular Sequence Data
Nanog Homeobox Protein
Promoter Regions, Genetic / genetics
Protein Binding
STAT Transcription Factors / genetics
STAT Transcription Factors / metabolism
Signal Transduction
Smad1 Protein / metabolism*
Stem Cells / cytology*
Stem Cells / drug effects
Stem Cells / metabolism*
T-Box Domain Proteins / genetics
T-Box Domain Proteins / metabolism

Substances

Bone Morphogenetic Proteins
DNA-Binding Proteins
Fetal Proteins
Homeodomain Proteins
Nanog Homeobox Protein
Nanog protein, mouse
STAT Transcription Factors
Smad1 Protein
Smad1 protein, mouse
T-Box Domain Proteins
Brachyury protein