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J Biol Chem, Vol. 273, Issue 46, 30460-30465, November 13, 1998

The Distal Regulatory Region of the Human Involucrin Promoter Is Required for Expression in Epidermis

James F. Crish, Tarif M. Zaim^§, and Richard L. Eckert^§¶

From the  Departments of Physiology and Biophysics, the ^§ Department of Dermatology, and the ^¶ Departments of Reproductive Biology, Biochemistry and Oncology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106-4970

Human involucrin (hINV) is a precursor of the keratinocyte cornified envelope that is specifically expressed in the suprabasal layers of stratifying squamous epithelia. The promoter distal (DRR) and proximal regulatory regions (PRR) are required for optimal in vitro expression (Welter, J. F., Crish, J. F., Agarwal, C., and Eckert, R. L. (1995) J. Biol. Chem. 270, 12614-12622; and Banks, E. B., Crish, J. F., Welter, J. F., and Eckert, R. L. (1998) Biochem. J. 331, 61-68). We now present the complete sequence of these regions and evaluate their ability to drive in vivo transcription. Transgenes containing 5000 or 2473 base pairs of upstream regulatory region drive tissue- and differentiation-appropriate expression in stratifying surface epithelia. In contrast, transgenes containing 1953, 1333, 986, or 41 base pairs of upstream regulatory region are not expressed in surface epithelia, indicating that loss of the DRR (nucleotides 2474/1953) results in loss of expression. Fusing the isolated DRR region directly to the hINV minimal promoter restores surface epithelial expression. Sequences downstream of the transcribed gene are not required for appropriate expression. The 1953/41 segment influences the pattern of differentiation-dependent expression. The 986/41 region, which includes the PRR, drives expression in internal epithelia.

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