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Originally published In Press as doi:10.1074/jbc.M209349200 on October 4, 2002

J. Biol. Chem., Vol. 277, Issue 49, 47190-47196, December 6, 2002
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Novel Target Sequences for Pax-6 in the Brain-specific Activating Regions of the Rat Aldolase C Gene*

Henriette Skala-Rubinson, Joëlle VinhDagger , Valérie LabasDagger , Axel Kahn, and Françoise Phan Dinh Tuy§

From the Département de Génétique, Développement et Pathologie Moléculaire, Institut Cochin, INSERM, CNRS, Université René Descartes, 24, rue du faubourg Saint Jacques, 75014 Paris and Dagger  Neurobiologie et Diversité Cellulaire, CNRS, Ecole Supérieure de Physique et de Chimie Industrielles, 10 rue Vauquelin, 75231 Paris, cedex 05, France

Upstream activating sequences of the rat aldolase C gene are shown here to confer brain-specific expression in transgenic mice. In addition to binding sites described previously for the brain-expressed POU proteins Brn-1 and Brn-2 (Skala, H., Porteu, A., Thomas, M., Szajnert, M. F., Okazawa, H., Kahn, A., and Phan-Dinh-Tuy, F. (1998) J. Biol. Chem. 273, 31806-31814), we have identified two novel DNA elements critical for an interaction with a brain-specific, high affinity DNA-binding protein. Characterization of this binding protein showed it to be sensitive to thiol oxidation and stable to heat at 100 °C. This protein was purified on the basis of its thermostability and its selective adsorption to streptavidin magnetic particles via a biotinylated multimer of its target DNA binding site. Liquid chromatography coupled to tandem mass spectrometry analysis, binding competition with consensus oligonucleotides, and antibody supershift assays led to its identification as the homeodomain paired protein Pax-6. This result suggests that the brain-specific aldolase C gene could constitute a new target for the transcription factor Pax-6, which is implicated increasingly in neurogenesis.

* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ To whom correspondence should be addressed. Tel.: 33-1-44-41-24-12; Fax: 33-1-44-41-24-21; E-mail: tuy@cochin.inserm.fr.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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