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J. Biol. Chem., Vol. 279, Issue 12, 11088-11095, March 19, 2004

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Mafs, Prox1, and Pax6 Can Regulate Chicken B1-Crystallin Gene Expression^*

Wenwu Cui, Stanislav I. Tomarev¶, Joram Piatigorsky¶, Ana B. Chepelinsky¶, and Melinda K. Duncan||

From the Department of Biological Sciences, University of Delaware, Newark, Delaware 19716 and the ^¶Laboratory of Molecular and Developmental Biology, National Eye Institute, National Institutes of Health, Bethesda, Maryland 20892-2730

During lens fiber cell differentiation, the regulation of crystallin gene expression is coupled with dramatic morphological changes. Here we report that Mafs, Prox1, and Pax6, which are essential transcription factors for normal lens development, bind to three functionally important cis elements, PL1, PL2, and OL2, in the chicken B1-crystallin promoter and may cooperatively direct the transcription of this lens fiber cell preferred gene. Gel shift assays demonstrated that Mafs bind to the MARE-like sequences in the PL1 and PL2 elements, whereas Prox1, a sequence-specific DNA-binding protein like its Drosophila homolog Prospero, interacts with the OL2 element. Furthermore, Pax6, a known repressor of the chicken B1-crystallin promoter, binds to all three of these cis elements. In transfection assays, Mafs and Prox1 activated the chicken B1-crystallin promoter; however, their transactivation ability was repressed when co-transfected with Pax6. Taken together with the known spatiotemporal expression patterns of Mafs, Prox1, and Pax6 in the developing lens, we propose that Pax6 occupies and represses the chicken B1-crystallin promoter in lens epithelial cells, and is displaced by Prox1 and Mafs, which activate the promoter, in differentiating cortical fiber cells.

Received for publication, November 12, 2003 , and in revised form, December 8, 2003.

^* This work was supported in part by National Eye Institute Grant EY12221 (to M. K. D.) and a Sigma Xi/NAS grant-in-aid (to W. C.) 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.

Supported by a Competitive Fellowship awarded by the University of Delaware Graduate School.

^|| To whom all correspondence should be addressed: Dept. of Biological Sciences, University of Delaware, Newark, DE 19716. Tel.: 302-831-0533; Fax: 302-831-2281; E-mail: duncanm{at}udel.edu.

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