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doi: 10.1242/10.1242/dev.00414

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Joint regulation of the MAP1B promoter by HNF3ß/Foxa2 and Engrailed is the result of a highly conserved mechanism for direct interaction of homeoproteins and Fox transcription factors

¹ CNRS UMR 8542, Ecole Normale Supérieure, 46 rue d'Ulm, 75230 Paris Cedex 05, France
² Université Paris 7, UFR de Biologie, 2 place Jussieu, 75005 Paris, France

^* Author for correspondence (e-mail: prochian{at}wotan.ens.fr)

Accepted 24 January 2003

The MAP1B (Mtap1b) promoter presents two evolutionary conserved overlapping homeoproteins and Hepatocyte nuclear factor 3ß (HNF3ß/Foxa2) cognate binding sites (defining putative homeoprotein/Fox sites, HF1 and HF2). Accordingly, the promoter domain containing HF1 and HF2 is recognized by cerebellum nuclear extracts containing Engrailed and Foxa2 and has regulatory functions in primary cultures of embryonic mesmetencephalic nerve cells. Transfection experiments further demonstrate that Engrailed and Foxa2 interact physiologically in a dose-dependent manner: Foxa2 antagonizes the Engrailed-driven regulation of the MAP1B promoter, and vice versa. This led us to investigate if Engrailed and Foxa2 interact directly. Direct interaction was confirmed by pull-down experiments, and the regions participating in this interaction were identified. In Foxa2 the interacting domain is the Forkhead box DNA-binding domain. In Engrailed, two independent interacting domains exist: the homeodomain and a region that includes the Pbx-binding domain. Finally, Foxa2 not only binds Engrailed but also Lim1, Gsc and Hoxa5 homeoproteins and in the four cases Foxa2 binds at least the homeodomain. Based on the involvement of conserved domains in both classes of proteins, it is proposed that the interaction between Forkhead box transcription factors and homeoproteins is a general phenomenon.

Key words: Cerebellum, Mes-metencephalon, Cytoskeleton, Homeodomain, Homeoprotein co-factors, Winged-helix/Forkhead box, Otx2, Goosecoid, Lim1

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