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First published online November 3, 2003
doi: 10.1242/10.1242/dev.00834
1 Department of Cell and Molecular Biology, Karolinska Institute, SE-171 77
Stockholm, Sweden
2 Department of Neuroscience, Medical Nobel Institute, Karolinska Institute,
SE-171 77 Stockholm, Sweden
* Author for correspondence (e-mail: Urban.Lendahl{at}cmb.ki.se)
Accepted 28 August 2003
The bone morphogenetic protein (BMP) and Notch signaling pathways are
crucial for cellular differentiation. In many cases, the two pathways act
similarly; for example, to inhibit myogenic differentiation. It is not known
whether this inhibition is caused by distinct mechanisms or by an interplay
between Notch and BMP signaling. Here we demonstrate that functional Notch
signaling is required for BMP4-mediated block of differentiation of muscle
stem cells, i.e. satellite cells and the myogenic cell line C2C12. Addition of
BMP4 during induction of differentiation dramatically reduced the number of
differentiated satellite and C2C12 cells. Differentiation was substantially
restored in BMP4-treated cultures by blocking Notch signaling using either the
-secretase inhibitor L-685,458 or by introduction of a
dominant-negative version of the Notch signal mediator CSL. BMP4 addition to
C2C12 cells increased transcription of two immediate Notch responsive genes,
Hes1 and Hey1, an effect that was abrogated by L-685,458. A
3 kb Hey1-promoter reporter construct was synergistically activated
by the Notch 1 intracellular domain (Notch 1 ICD) and BMP4. The BMP4 mediator
SMAD1 mimicked BMP activation of the Hey1 promoter. A synthetic
Notch-responsive promoter containing no SMAD1 binding sites responded to
SMAD1, indicating that DNA-binding activity of SMAD1 is not required for
activation. Accordingly, Notch 1 ICD and SMAD1 interacted in binding
experiments in vitro. Thus, the data presented here provide evidence for a
direct interaction between the Notch and BMP signaling pathways, and indicate
that Notch has a crucial role in the execution of certain aspects of
BMP-mediated differentiation control.
Key words: SMAD, Delta, Serrate, TGFß, skeletal muscle,
-secretase inhibitor
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