A functional analysis of ACP-20, an adult-specific cuticular protein gene from the beetle Tenebrio: role of an intronic sequence in transcriptional activation during the late metamorphic period

A gene encoding the adult cuticular protein ACP‐20 was isolated in Tenebrio. It consists of three exons interspersed by two introns, intron 1 interrupting the signal peptide. To understand the regulatory mechanisms of ACP‐20 expression, ACP‐20 promoter–luciferase reporter gene constructs were transf...

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Bibliographic Details
Published in:Insect molecular biology 2004-10, Vol.13 (5), p.481-493
Main Authors: Lemoine, A., Mathelin, J., Braquart-Varnier, C., Everaerts, C., Delachambre, J.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Base Sequence
Biodiversity and Ecology
cotransfection
Ecdysone - metabolism
Ecdysone - pharmacology
ecdysteroids
Environmental Sciences
Epidermis - metabolism
Gene Library
Genes, Reporter - physiology
Genetics
insect cuticular protein gene regulation
Insect Proteins - genetics
Insect Proteins - physiology
intron
Introns - genetics
Introns - physiology
Life Sciences
Luciferases - metabolism
metamorphosis
Metamorphosis, Biological - genetics
Metamorphosis, Biological - physiology
Molecular Sequence Data
Populations and Evolution
Promoter Regions, Genetic - drug effects
Promoter Regions, Genetic - physiology
Sequence Analysis, DNA
Tenebrio
Tenebrio - genetics
Tenebrio - physiology
Tenebrionidae
Transcriptional Activation - genetics
Transcriptional Activation - physiology
Transfection
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Summary:A gene encoding the adult cuticular protein ACP‐20 was isolated in Tenebrio. It consists of three exons interspersed by two introns, intron 1 interrupting the signal peptide. To understand the regulatory mechanisms of ACP‐20 expression, ACP‐20 promoter–luciferase reporter gene constructs were transfected into cultured pharate adult wing epidermis. Transfection assays needed the presence of 20‐hydroxyecdysone, confirming that ACP‐20 is up‐regulated by ecdysteroids. Analysis of 5′ deletion constructs revealed that three regions are necessary for high levels of transcription. Interaction experiments between intronic fragments and epidermal nuclear proteins confirmed the importance of intron 1 in ACP‐20 transcriptional control, which results from the combined activity of regulatory cis‐acting elements of the promoter and those of intron 1.
ISSN:0962-1075
1365-2583
DOI:10.1111/j.0962-1075.2004.00508.x