8-Br-cAMP inhibits the transient expression of firefly luciferase

G. T. Coker, L. Vinnedge, K. L. O'Malley

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

The genes for firefly luciferase and chloramphenicol acetyltransferase (CAT) were used as reporter genes to explore the activation of heterologous promoters by 8-Br-cAMP. Cells were transfected with a CAT gene/tyrosine hydroxylase promoter, which contains a cAMP response element. Extracts from cells treated with 8-Br-cAMP had 340% more enzyme activity than untreated cells. In contrast, treated cells transfected with a tyrosine hydroxylase/luciferase construct had 30% less activity than control cells. Simian virus and rous sarcoma virus promoters/luciferase constructs also had lower activities in cells treated with 8-Br-cAMP than untreated cells. The inhibition of luciferase enzyme activity by cAMP appears to be posttranscriptional since both luciferase and CAT RNA levels were similarly increased in cells treated with 8-Br-cAMP or 1-methyl-3-isobutylmethylxanthine. The lower level of luciferase activity was not due to simple allosteric inhibition. We conclude that constructs using the firefly luciferase as a reporter gene are unsuitable for studying the effects of cAMP on the regulation of promoters.

Original languageEnglish
Pages (from-to)183-185
Number of pages3
JournalFEBS Letters
Volume249
Issue number2
DOIs
StatePublished - Jun 5 1989

Keywords

  • Bromo cyclic AMP, 8-
  • Luciferase
  • Methyl-3-isobutylmethylxanthine, 1-
  • Reporter gene
  • Transfection

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