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 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.
Lemoine
A
A
UMR CNRS 5548, Développement et Communication Chimique chez les Insectes, Université de , Dijon, France. aleth.lemoine@u-bourgogne.fr
Mathelin
J
J
Braquart-Varnier
C
C
Everaerts
C
C
Delachambre
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004440:Ecdysone / Q000378:metabolism / Q000494:pharmacology; D004817:Epidermis / Q000378:metabolism; D015723:Gene Library; D017930:Genes, Reporter / Q000502:physiology; D019476:Insect Proteins / Q000235:genetics* / Q000502:physiology*; D007438:Introns / Q000235:genetics / Q000502:physiology; D008156:Luciferases / Q000378:metabolism; D008675:Metamorphosis, Biological / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000502:physiology; D017422:Sequence Analysis, DNA; D013712:Tenebrio / Q000235:genetics* / Q000502:physiology; D015533:Transcriptional Activation / Q000235:genetics / Q000502:physiology*; D014162:Transfection
D019476:Insect Proteins; C075200:ACP-20 protein, Tenebrio molitor; D004440:Ecdysone; D008156:Luciferases