Regulation of fos-lacZ fusion gene expression in primary mouse epidermal keratinocytes isolated from transgenic mice

Wendy B Bollag, Y. Xiong, J. Ducote, C. S. Harmon

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The expression of a fos-lacZ fusion gene was studied in primary mouse epidermal keratinocytes obtained from transgenic mice. This gene construct contains the entire upstream regulatory sequence of c-fos, and expression of the endogenous and fusion gene was shown by Northern analysis to correlate upon induction with the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA). Using a chromogenic substrate of β-galactosidase, we also demonstrated that expression of the fusion gene product, like that of Fos, was localized to the cell nucleus. In addition, we showed that epidermal keratinocytes responded to dialysed fetal bovine serum (FBS), TPA and high-calcium medium with enhanced Fos-lacZ expression and an inhibition of proliferation. The time course of induction of Fos-lacZ expression was similar for dialysed FBS and TPA, with a peak ~2 h after exposure. Exposure for ~24 h to an elevated extracellular calcium concentration was required to elicit an increase in Fos-lacZ expression. The lack of an immediate effect of raising medium calcium levels on Fos-lacZ expression contrasted with the rapidity of its effect on DNA synthesis, which was significantly inhibited within 6-8 h. In addition, we found that the protein kinase C inhibitor Ro 31-7549 blocked Fos-lacZ expression induced by TPA but had little or no effect on that elicited by high calcium levels. Thus, although our results indicate that the fos gene product may be involved in mediating epidermal keratinocyte growth arrest in response to differentiative agents such as FBS, TPA and high medium calcium levels, the exact role of this gene product remains unclear.

Original languageEnglish (US)
Pages (from-to)263-270
Number of pages8
JournalBiochemical Journal
Volume300
Issue number1
DOIs
StatePublished - Jan 1 1994

Fingerprint

Lac Operon
Tetradecanoylphorbol Acetate
Keratinocytes
Gene expression
Transgenic Mice
Fusion reactions
Genes
Calcium
Acetates
Gene Expression
Gene Fusion
Serum
Galactosidases
fos Genes
Chromogenic Compounds
Protein C Inhibitor
Phorbol Esters
Protein Kinase Inhibitors
Cell Nucleus
Protein Kinase C

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Regulation of fos-lacZ fusion gene expression in primary mouse epidermal keratinocytes isolated from transgenic mice. / Bollag, Wendy B; Xiong, Y.; Ducote, J.; Harmon, C. S.

In: Biochemical Journal, Vol. 300, No. 1, 01.01.1994, p. 263-270.

Research output: Contribution to journalArticle

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