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J Biol Chem, Vol. 273, Issue 50, 33848-33855, December 11, 1998

Cloning and Characterization of the Human ₄-Integrin Gene Promoter and Enhancers

Asako Suzuki Takaoka^§, Tesshi Yamada, Masahiro Gotoh, Yae Kanai, Kohzoh Imai^§, and Setsuo Hirohashi

From the  Pathology Division, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045 and the ^§ First Department of Internal Medicine, Sapporo Medical University School of Medicine, Minami-1, Nishi-16, Chuo-ku, Sapporo 060-8543, Japan

The cell-surface adhesion molecule ₆₄-integrin is a receptor for laminins and a component of hemidesmosomes. ₄-Integrin expression is restricted to proliferating basal keratinocytes in the epidermis and is suppressed when differentiation commences. Altered ₄-integrin expression levels correlate significantly with the aggressive behavior of cancers. In order to clarify the mechanisms that regulate transcription of the ₄-integrin gene, we cloned its 5'-flanking region. This 5'-flanking region was found to have a high G + C content and not to contain either TATA or CAAT boxes. Nested delimitation and reporter analyses mapped a basal promoter to nucleotides 106 to +105, surrounding the most proximal transcription initiation site. Gel retardation and mutational analyses revealed that cooperation between AP1 and Ets, interacting with other factors, mediated the promoter activity. In addition to the promoter element, enhancer activity was found in the first intron (+1905/+3933) and in a sequence upstream of the promoter region (414/107). These findings should facilitate our understanding of the regulation of ₄-integrin gene expression in processes such as cell growth and differentiation, apoptosis, and cancer development and metastasis.

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