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Nucleic Acids Research, 2001, Vol. 29, No. 3 732-742
© 2001 Oxford University Press

Smad proteins function as co-modulators for MEF2 transcriptional regulatory proteins

Zoë A. Quinn1,2, Chih-Chin Yang1, Jeffrey L. Wrana3 and John C. McDermott1,2,*

1Department of Biology, York University, Toronto, Ontario M3J 1P3, Canada, 2Department of Biological Sciences, University of Durham, Durham DH1 3LE, UK and 3Program in Molecular Biology and Cancer, Samuel Lunenfield Research Institute, Mount Sinai Hospital, Room 1075, 600 University Avenue, University of Toronto, Toronto, Ontario M5G 1X5, Canada

An emerging theme in transforming growth factor-ß (TGF-ß) signalling is the association of the Smad proteins with diverse groups of transcriptional regulatory proteins. Several Smad cofactors have been identified to date but the diversity of TGF-ß effects on gene transcription suggests that interactions with other co-regulators must occur. In these studies we addressed the possible interaction of Smad proteins with the myocyte enhancer-binding factor 2 (MEF2) transcriptional regulators. Our studies indicate that Smad2 and 4 (Smad2/4) complexes cooperate with MEF2 regulatory proteins in a GAL4-based one-hybrid reporter gene assay. We have also observed in vivo interactions between Smad2 and MEF2A using co-immunoprecipitation assays. This interaction is confirmed by glutathione S-transferase pull-down analysis. Immunofluorescence studies in C2C12 myotubes show that Smad2 and MEF2A co-localise in the nucleus of multinuclear myotubes during differentiation. Interestingly, phospho-acceptor site mutations of MEF2 that render it unresponsive to p38 MAP kinase signalling abrogate the cooperativity with the Smads suggesting that p38 MAP Kinase-catalysed phosphorylation of MEF2 is a prerequisite for the Smad–MEF2 interaction. Thus, the association between Smad2 and MEF2A may subserve a physical link between TGF-ß signalling and a diverse array of genes controlled by the MEF2 cis element.

* To whom correspondence should be addressed at: 327 Farquharson, LSB, York University, 4700 Keele Street, Toronto, Ontario M3J 1P3, Canada. Tel: +1 416 736 2100; Fax: +1 416 736 5698; Email: jmcderm{at}yorku.ca


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