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Nucleic Acids Research, 1990, Vol. 18, No. 2 247-253
© 1990

MOLECULAR BIOLOGY

Epithelial-specific keratin gene expression: identification of a 300 base-pair controlling segment

C.-K. Jiang1,+, H.S. Epstein1, M. Tomic§,1, I.M. Freedberg1 and M. Blumenberg1,2,*

1Departments of Dermatology, NYU Medical Center 550 First Ave., New York, NY 10016, USA 2Departments of Biochemistry, NYU Medical Center 550 First Ave., New York, NY 10016, USA

* To whom correspondence should be addressed

Received October 16, 1989. Revised December 5, 1989. Accepted December 5, 1989.

To elucidate the elements required for regulation of keratin expression in epidermis, we have linked a short, 300 base pair segment, corresponding to the promoter region of a human K # 14 gene, to the chloramphenicolacetyl-transferase gene. This construct was introduced into various mammalian cell lines and primary cultures via Ca3(PO4)2 precipitation. The 300 base pair segment from the keratin gene promoter region was active in all epithelial cells studied including transformed, simple epithelial cells such as HeLa and ME-180, cell lines derived from stratified epithelium, such as SCC-12, as well as primary cultures of epithelial cells. The construct was inactive in all non-epithelial cells tested including fibroblasts and melanocytes. The segment does not function as a silencer in nonepithelial cells but it can function as an enhancer in epithelial cells. Using the polymerase chain reaction we have constructed a series of deletions of the promoter and have localized an essential function within a 40 bp sequence. We conclude that we have identified the keratin gene promoter that is sufficient to confer epithelial-specific expression.

Present address: +Shanghai Institute of Cell Biology, Academia Sinica, 320 Yo-Yand Road, Shanghai, China

Present address: §Institute Boris Kidric, Vinca, Yugoslavia


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