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Development 129, 2195-2207 (2002)
© 2002 The Company of Biologists Limited

Hes6 regulates myogenic differentiation

Judy Cossins1, Ann E. Vernon2,*, Yun Zhang1,*, Anna Philpott2 and Philip H. Jones2,{dagger}

1 Cancer Research UK, Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, Oxford, UK
2 Cancer Research UK, Department of Oncology, University of Cambridge, Hutchinson/MRC Research Centre, Addenbrooke’s Hospital, Cambridge, UK
* These authors contributed equally to this work

{dagger}Author for correspondence (e-mail: phj20{at}cam.ac.uk)

Accepted 12 February 2002

Hes6 is a basic helix-loop-helix transcription factor homologous to Drosophila Enhancer of Split (EoS) proteins. It is known to promote neural differentiation and to bind to Hes1, a related protein that is part of the Notch signalling pathway, affecting Hes1-regulated transcription. We show that Hes6 is expressed in the murine embryonic myotome and is induced on C2C12 myoblast differentiation in vitro. Hes6 binds DNA containing the Enhancer of Split E box (ESE) motif, the preferred binding site of Drosophila EoS proteins, and represses transcription of an ESE box reporter. When overexpressed in C2C12 cells, Hes6 impairs normal differentiation, causing a decrease in the induction of the cyclin-dependent kinase inhibitor, p21Cip1, and an increase in the number of cells that can be recruited back into the cell cycle after differentiation in culture. In Xenopus embryos, Hes6 is co-expressed with MyoD in early myogenic development. Microinjection of Hes6 RNA in vivo in Xenopus embryos results in an expansion of the myotome, but suppression of terminal muscle differentiation and disruption of somite formation at the tailbud stage. Analysis of Hes6 mutants indicates that the DNA-binding activity of Hes6 is not essential for its myogenic phenotype, but that protein-protein interactions are. Thus, we demonstrate a novel role for Hes6 in multiple stages of muscle formation.

Key words: Muscle, Notch, Enhancer of split, Mouse, Xenopus




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© The Company of Biologists Ltd 2002