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(Investigative Ophthalmology and Visual Science. 2004;45:1080-1087.)
© 2004 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.03-1180
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Regulation of Involucrin Expression in Normal Human Corneal Epithelial Cells: A Role for Activator Protein One

Gautam Adhikary,1 James Crish,1 Jonathan Lass,2 and Richard L. Eckert1,3,4,5

1From the Departments of Physiology and Biophysics, 2Ophthalmology, 3Oncology, 4Dermatology, and 5Biochemistry, Case School of Medicine, Cleveland, Ohio.

PURPOSE. Understanding the mechanisms that regulate gene expression in the human cornea is an important goal. In the present study, the involucrin gene was used as a model to study this regulation. Human involucrin (hINV) is a structural protein that is selectively expressed in surface epithelia, including corneal epithelial cells.

METHODS. Regulation of involucrin gene expression was monitored in cultures of normal human primary corneal epithelial cells.

RESULTS. The studies revealed that an activator protein (AP)-1 DNA-binding site is essential for appropriate basal and stimulus-dependent hINV promoter activity. Mutation of this site, AP1-1, results in a loss of hINV promoter activity. A gel mobility supershift analysis revealed interaction of the AP1 factors, Fra-1, Fra-2, and JunB, with this element. Inhibition of AP1 function with a dominant-negative form of AP1 also inhibited expression. Treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA), a protein kinase C activator, increased hINV gene expression, a response that correlates with increased nuclear AP1 factor level and binding to the hINV gene AP1-1 response element. Expression of the endogenous hINV gene is also increased by TPA treatment.

CONCLUSIONS. These findings point to an important role for AP1 transcription factors in the regulation of human corneal epithelial cell involucrin gene expression.




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