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J Biol Chem, Vol. 273, Issue 48, 31939-31946, November 27, 1998

Intestinal Expression of the Calbindin-D9K Gene in Transgenic Mice
REQUIREMENT FOR A Cdx2-BINDING SITE IN A DISTAL ACTIVATOR REGION

Sabine Colnot, Béatrice Romagnolo^¶, Mireille Lambert, Françoise Cluzeaud^**, Arlette Porteu^¶, Alain Vandewalle^**, Monique Thomasset, Axel Kahn^¶, and Christine Perret^¶

From  INSERM U458, Hôpital Robert Debré, 48 boulevard Sérurier, 75019 Paris, France, ^¶ INSERM U129, Institut Cochin de Génétique Moléculaire, 24 rue du faubourg St Jacques, 75014 Paris, France, and ^** INSERM U246, Faculté de Médecine Xavier Bichat, Institut Fédératif de Recherche 02, B.P. 416, 75810 Paris Cedex 18, France

The calbindin-D9K gene encodes a vitamin D-induced calcium-binding protein that is expressed as a marker of small intestine differentiation. We have shown that 4580 base pairs of its 5' DNA regulatory region can target reporter transgene expression in the intestine and cause this transgene to respond like the endogenous gene to vitamin D active metabolite and that the homeoprotein Cdx2 is bound to the TATA box in the intestine. We now show that the 4580 base pairs construct confers a differentiated pattern of reporter transgene expression in the intestine and that cooperation between the proximal promoter and a distal element located in an opened chromatin structure is responsible for the intestinal expression and vitamin D responsiveness of the transgene. Gel shift and footprinting assays using duodenal nuclear extracts indicate that this distal element contains a Cdx2-binding site. Finally, a mutation in this distal Cdx2-binding site dramatically decreases intestinal expression in transgenic mice. This report, using an in vivo approach, demonstrates the crucial role of Cdx2 for the transcription of an intestinal gene.

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