Upregulation of the Drosophila 1731 retrotransposon long-terminal repeat by UV-B irradiation requires a short sequence in the U3 region.
A 1731 is a Drosophila melanogaster retrotransposon, the transcripts of which decrease in Drosophila cells after treatment by the 20-hydroxyecdysone (20-OH), the steroid-molting hormone of insects. In order to analyze the regulation of the long terminal repeat (LTR) directed transcription by UV-B, S2 Drosophila cells were transfected with various chimeric constructs carrying the LTR of 1731 linked to the bacterial chloramphenicol acetyltransferase (CAT) reporter gene and then subjected to UV-B irradiation. The results demonstrated that the 1731 LTR is activated by UV-B irradiation in a dose- and time-dependent manner. Using constructions expressing the reporter gene under the control of either the entire or deleted LTRs of 1731, we established that a sequence located in the U3 region was required for the retrotransposon to respond to UV-B. The cis-acting element is identical to the binding sequence of the dorsal transcription factors. In addition, the S2 Drosophila cell produced extracellular factor(s) in response to UV-B irradiation which activated the 1731 LTR in nonirradiated cells. This factor(s) was detected when responding cells were cocultured with inducing cells and when conditioned medium from irradiated cultures was added to the cell cultures.
Faure
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Institut de Chimie Biologique, Université de Provence, Marseille, France.
Best-Belpomme
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Champion
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D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004247:DNA / Q000235:genetics / Q000528:radiation effects; D004331:Drosophila melanogaster / Q000235:genetics* / Q000528:radiation effects / Q000821:virology; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D012091:Repetitive Sequences, Nucleic Acid / Q000528:radiation effects*; D018626:Retroelements / Q000528:radiation effects*; D015398:Signal Transduction; D014162:Transfection; D014466:Ultraviolet Rays; D015854:Up-Regulation
D018626:Retroelements; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase