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J. Biol. Chem., Vol. 279, Issue 10, 9103-9114, March 5, 2004

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Krüppel-like Factors Regulate the Lama1 Gene Encoding the Laminin 1 Chain^*

Silvia A. Piccinni, Anne-Laure Bolcato-Bellemin¶, Annick Klein, Vincent W. Yang||, Michèle Kedinger, Patricia Simon-Assmann, and Olivier Lefebvre**

From the INSERM Unit 381, Development and Pathology of the Digestive Tract, University Louis Pasteur, 67 200 Strasbourg, France and ^||Emory University School of Medicine, Division of Digestive Diseases, Atlanta, Georgia 30322

Laminin-1 (111), a basement membrane (BM) constituent, has been associated with differentiation processes and also with malignant progression. In the intestinal tissue, the 1 chain is expressed and secreted in the subepithelial BM during the developmental period; in the adult rodent tissue, it is restricted to the BM of the dividing cells. To understand how laminin 1 chain expression is regulated, we cloned and characterized a 2-kb promoter region of the Lama1 mouse gene. Analysis of the promoter was conducted in the Caco2-TC7 intestinal epithelial cells by transient transfection of serially deleted and site-directed mutated promoter constructs, by electrophoretic mobility shift assays, and expression of selected transcription factors. We determined that a proximal region, which includes an Sp1-binding GC box and a Krüppel-like element, was important for the promoter activity. This region is conserved between the human and mouse genes. Interestingly, two Krüppel-like factors KLF4 and KLF5 exhibit opposing effects on the Lama1 promoter activity that are decreased and increased, respectively, in the intestinal epithelial cells. These data corroborate the complementary expression of KLF4 and KLF5 along the intestinal crypt-villus axis and the parallel expression of KLF5 and laminin 1 chain in the crypt region. Finally, we showed that glucocorticoids stimulate the promoter activity. This study is the first characterization of the Lama1 promoter; we identified regulatory elements that may account for the expression pattern of the endogenous protein in the mouse intestine.

Received for publication, June 3, 2003 , and in revised form, November 6, 2003.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AJ519495.

^* This work was supported in part by grants from INSERM, the Research Ministry (ACI Developmental Biology and Physiopathology 172), and the Ligue Contre le Cancer, Comité Départemental du Haut-Rhin. 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.

Both authors contributed equally to this work.

Recipient of a fellowship from the French Ministry of Research and Education.

^¶ Recipient of a fellowship from the Association de la Recherche Contre le Cancer.

^** To whom correspondence should be addressed: INSERM-Unité 381, 3, Ave. Molière, 67 200 Strasbourg, France. Tel.: 33-3-88-27-77-27; Fax: 33-3-88-26-35-38; E-mail: olivier.lefebvre{at}INSERM.u-strasbg.fr.

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