Identification of a strong transcriptional activator for the copia retrotransposon responsible for its differential expression in Drosophila hydei and melanogaster cell lines

Biochem Biophys Res Commun. 1994 Aug 30;203(1):392-9. doi: 10.1006/bbrc.1994.2195.

Abstract

We have characterized the regulatory properties of a 72bp sequence located in the 5' untranslated domain of the Drosophila copia retrotransposon, 3' to the left LTR, by transient transfection assays with cell lines derived from either Drosophila hydei (DH33 cells) or Drosophila melanogaster (Schneider II and Kc cells). Reporter plasmids were constructed which contained the lacZ gene under the control of either the entire copia LTR with 5' untranslated domain, or a minimal heterologous promoter flanked with the identified copia regulatory sequences. Upon transfection into the copia-free DH33 cells, the presence of the 72bp sequence resulted for all reporter plasmids in a 100-700 fold increase in expression level -as well as in reporter gene RNA levels- whereas this sequence had no enhancing effect upon transfection of the same plasmids into the copia-containing Schneider II or Kc cells. Moreover, mobility shift assays with the 72bp enhancer sequence disclosed two specific bands of retarded mobility with whole-cell extracts from DH33 cells, whereas no retarded band could be detected, under identical conditions, with extracts from Schneider II cells. UV crosslinking experiments between the enhancer sequence and DH33 extracts revealed a single protein species -of app. mol. wt. 50kD- for both retarded bands, thus strongly suggesting that they simply correspond to the sequential binding of two identical factor molecules to the enhancer sequence. These data demonstrate that the copia-free D. hydei cells express a strong transcriptional activator for the copia element and possible interpretations for the absence of this factor in the copia-containing D. melanogaster cells are discussed in terms of a possible "adaptation" of the "host" (D. melanogaster) to an otherwise highly mutagenic "parasite" (copia with its transcription factor).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites
  • Cell Line
  • Consensus Sequence
  • DNA Primers
  • DNA Transposable Elements*
  • Drosophila / genetics*
  • Drosophila melanogaster / genetics*
  • Enhancer Elements, Genetic
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Repetitive Sequences, Nucleic Acid
  • Retroviridae
  • Transcription Factors / isolation & purification
  • Transcription Factors / metabolism*
  • Transcription, Genetic*
  • Transfection
  • beta-Galactosidase / biosynthesis

Substances

  • DNA Primers
  • DNA Transposable Elements
  • Transcription Factors
  • beta-Galactosidase