Regulation of human profilaggrin promoter activity in cultured epithelial cells by retinoic acid and glucocorticoids

Richard B. Presland, Marjana Tomic-Canic, S. Patrick Lewis, Beverly A. Dale

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Vitamin A and other retinoids profoundly inhibit both morphological and biochemical aspects of epidermal differentiation in vitro. Profilaggrin, like most other markers of keratinocyte differentiation, is negatively regulated by retinoic acid in vitro, both at the level of mRNA synthesis and by inhibiting the activity of endoproteases that convert profilaggrin to filaggrin. Profilaggrin is an abundant component of keratohyalin granules and forms the precursor of filaggrin, the keratin associated protein of the stratum corneum. In this report, we identify a region of the human profilaggrin promoter that is involved in the transcriptional regulation of expression by retinoic acid (RA). A series of promoter deletions linked to the chloramphenicol acetyl transferase (CAT) reporter gene were prepared and analyzed by transfection into Hela cells and keratinocytes. We also cotransfected vectors expressing retinoic acid receptor and cultured the transfected cells in the presence and absence of ligand. The region responsive to retinoic acid was localized to a 53 bp sequence between - 1109 and - 1056 (relative to the mRNA start site at + 1) that contains a cluster of five retinoic acid response elements with variable spacing and orientation. In vitro gel shift analysis demonstrated that nuclear retinoid receptors do not bind directly to the identified sequence, suggesting that the mode of regulation by RA may be indirect or that binding requires another cofactor in addition to retinoid receptors. Whereas in keratin genes retinoic acid and glucocorticoid responsive sequences frequently coincide, the glucocorticoid response element in the profilaggrin promoter was located downstream of the RARE cluster between - 965 and - 951. These studies demonstrate that RA and glucocorticoids regulate profilaggrin expression at least in part by transcriptional mechanisms, via a region of the promoter that contains both retinoid and glucocorticoid responsive elements.

Original languageEnglish
Pages (from-to)192-205
Number of pages14
JournalJournal of Dermatological Science
Volume27
Issue number3
DOIs
StatePublished - Oct 24 2001
Externally publishedYes

Fingerprint

Tretinoin
Glucocorticoids
Cultured Cells
Epithelial Cells
Retinoids
Response Elements
Keratins
Keratinocytes
Genes
Messenger RNA
Retinoic Acid Receptors
filaggrin
Differentiation Antigens
Electrophoretic Mobility Shift Assay
Chloramphenicol
Cytoplasmic and Nuclear Receptors
Transferases
Vitamin A
Reporter Genes
HeLa Cells

Keywords

  • CAT assay
  • Glucocorticoid
  • Profilaggrin
  • Retinoic acid
  • Transfection

ASJC Scopus subject areas

  • Dermatology

Cite this

Regulation of human profilaggrin promoter activity in cultured epithelial cells by retinoic acid and glucocorticoids. / Presland, Richard B.; Tomic-Canic, Marjana; Lewis, S. Patrick; Dale, Beverly A.

In: Journal of Dermatological Science, Vol. 27, No. 3, 24.10.2001, p. 192-205.

Research output: Contribution to journalArticle

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