Effects of modulators of tyrosinase activity on expression of murine interleukin-2 cDNA driven by the tyrosinase promoter

N. Miller, Richard Geoffrey Vile, I. R. Hart

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Sequence analysis of the promoter region of the murine tyrosinase gene identified various consensus motifs including AP2 sites, cAMP and TPA response elements (CREs/TREs) and retinoic acid response element (RARE) half-sites. By linking two different promoter lengths (2.5 kb or 769 bp) to murine interleukin-2 (IL-2) cDNA we have used IL-2 production by transduced B16 cells to monitor response to inducing agents capable of acting through these elements. Aminophylline or theophylline (0.1-2 mM) added to the culture medium of transfected B16, but not 3T3, cells, increased IL-2 secretion significantly (P > 0.05) in a dose dependent fashion. This response was comparable In cells transfected with either the full length or the truncated promoter. Therefore, the cAMP responsiveness of the tyrosinase promoter probably is mediated by CREs and not AP2 sites, since the truncated promoter contains the former but not the latter regions. Retinoic acid at various concentrations (0.1-1 μM) evoked a standard increase in IL-2 production. Responses were similar for both promoter constructs, which suggests either that each RARE half-site can confer the full retinoic acid response, or that retinoic acid is mediating its effect through pathways independent of the RARE sites. TPA (2 nM-2 μM) had no effect on IL-2 production. These results demonstrate that the tyrosinase promoter can be induced by certain pharmacological agents and raise the possibility that administration of such substances may enhance expression of therapeutic genes controlled by this promoter.

Original languageEnglish (US)
Pages (from-to)75-81
Number of pages7
JournalMelanoma Research
Volume5
Issue number2
StatePublished - 1995
Externally publishedYes

Fingerprint

Monophenol Monooxygenase
Tretinoin
Interleukin-2
Complementary DNA
Response Elements
Aminophylline
3T3 Cells
Theophylline
Genetic Promoter Regions
Sequence Analysis
Culture Media
Pharmacology
Gene Expression
Genes

Keywords

  • Murine tyrosinase promoter
  • Regulation
  • Retinoic acid
  • Theophylline

ASJC Scopus subject areas

  • Cancer Research
  • Dermatology

Cite this

Effects of modulators of tyrosinase activity on expression of murine interleukin-2 cDNA driven by the tyrosinase promoter. / Miller, N.; Vile, Richard Geoffrey; Hart, I. R.

In: Melanoma Research, Vol. 5, No. 2, 1995, p. 75-81.

Research output: Contribution to journalArticle

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