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J. Biol. Chem., Vol. 279, Issue 23, 23874-23881, June 4, 2004

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Disruption of Meox or Gli Activity Ablates Skeletal Myogenesis in P19 Cells^*

Helen Petropoulos¶, Peter J. Gianakopoulos||, Alan G. Ridgeway**, and Ilona S. Skerjanc

From the Department of Biochemistry, Medical Sciences Building, The University of Western Ontario, London, Ontario N6A 5C1, Canada and the ^**Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115

Gli2 and Meox1 are transcription factors that are expressed in the developing somite and play roles in the commitment of cells to the skeletal muscle lineage. To further define their roles in regulating myogenesis, the function of wild type and dominant-negative forms of Gli2 and Meox1 were examined in the context of differentiating P19 stem cells. We found that Gli2 overexpression up-regulated transcript levels of Meox1 and, conversely, Meox1 overexpression resulted in the upregulation of Gli2 transcripts. Furthermore, dominant-negative forms of either Meox1 or Gli2 disrupted the ability of P19 cells to commit to the muscle lineage and to properly express either Gli2 or Meox1, respectively. Finally, Pax3 transcripts were induced by Gli2 overexpression and lost in the presence of either mutants Meox1 or Gli2. Taken together, these results support the existence of a regulatory loop between Gli2, Meox1, and Pax3 that is essential for specification of mesodermal cells into the muscle lineage.

Received for publication, November 18, 2003 , and in revised form, March 5, 2004.

^* This work was supported in part by Grant MOP-49605 (to I. S. S.) from the Neuromuscular Research Partnership, an initiative of ALS Canada, Muscular Dystrophy Association of Canada, and the Canadian Institutes of Health Research. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Both authors contributed equally to this manuscript.

^¶ Supported by a Natural Sciences and Engineering Research Council of Canada postgraduate studentship and an Ontario graduate scholarship.

^|| Supported by a Premier's Research Excellence Award in partnership with the Foundation for Gene and Cell Therapy.

Supported by a Canadian Institute of Aging Investigator Award. To whom correspondence should be addressed: Dept. of Biochemistry, Medical Sciences Building, University of Western Ontario, London, Ontario N6A 5C1, Canada. Tel.: 519-661-2111 (ext. 86867); Fax: 519-661-3175; E-mail: skerjanc{at}uwo.ca.

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