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8‐Br‐cAMP inhibits the transient expression of firefly luciferase
Author(s) -
Coker G.T.,
Vinnedge L.,
O'Malley K.L.
Publication year - 1989
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(89)80620-9
Subject(s) - luciferase , chloramphenicol acetyltransferase , rous sarcoma virus , reporter gene , promoter , microbiology and biotechnology , transfection , heterologous , tyrosine hydroxylase , chemistry , acetyltransferase , gene expression , biology , gene , enzyme , biochemistry , acetylation
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.