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J. Biol. Chem., Vol. 282, Issue 45, 33192-33200, November 9, 2007

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A Muscle-specific Promoter Directs Pitx3 Gene Expression in Skeletal Muscle Cells^*

Vincent Coulon¹², Aurore L'Honoré¹³, Jean-François Ouimette¹, Émilie Dumontier, Pepijn van den Munckhof^¶, and Jacques Drouin⁴

From the Laboratoire de Génétique Moléculaire, Institut de Recherches Cliniques de Montréal, Montréal, Quebec H2W 1R7, Canada, CNRS UMR 5535, Institut de Génétique Moléculaire, 1919 Route de Mende, 34293 Montpellier Cedex 05, France, and ^¶Academisch Medisch Centrum, Universiteit van Amsterdam, Meibergdreef 9, Posthus 22660, Amsterdam 1100DD, The Netherlands

The Pitx homeobox transcription factor genes have been implicated in different developmental processes, including determination of hind limb identity for Pitx1, left-right asymmetry for Pitx2, and eye development and survival of midbrain dopaminergic neurons for Pitx3. Pitx1 and Pitx2 have partly redundant activities in craniofacial development, including in pituitary organogenesis, as indicated by their names. These genes also exhibit redundant activities in the control of hind limb bud growth. Recent studies have shown expression of the three Pitx genes in muscle, with Pitx3 being the most widely expressed in all skeletal muscles. We now report the identification of a muscle-specific promoter within the Pitx3 gene that is situated between the first exon for eye and brain expression and exon 2 that contains the initiator ATG codon. Sequences proximal to this muscle-specific exon 1 are essential and sufficient to confer muscle-specific expression in transgenic mice, they are responsive to myogenic basic helix-loop-helix regulatory factors, and they recruit these factors in vivo. In agreement with exclusive use of the muscle-specific promoter in aphakia mice that are deleted of the brain promoter, the trimethyl-lysine 4 histone H3 promoter signature shifts to this promoter in embryonic day 13 ak limb bud muscle cells. Myogenic basic helix-loop-helix regulatory factor activation of Pitx3 transcription may be part of a positive feedback loop contributing to establishment of the myogenic program.

Received for publication, July 25, 2007 , and in revised form, September 4, 2007.

^* This work was supported in part by grants from 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.

¹ These authors contributed equally to this work.

² Supported by a fellowship from INSERM and by a Lavoisier fellowship from the French Ministère des AffairesÉtrangères.

³ Supported by the Fondation pour la Recherche Médicale.

⁴ To whom correspondence should be addressed: Laboratoire de Génétique Moléculaire, Institut de Recherches Cliniques de Montréal, 110 Ave. des Pins Ouest, Montréal, Quebec H2W 1R7, Canada. Tel.: 514-987-5680; Fax: 514-987-5575; E-mail: jacques.drouin{at}ircm.qc.ca.

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