Characterization of the reverse transcriptase of 1731, a Drosophila melanogaster retrotransposon.
The nucleotide sequence of 1731, a retrotransposon cloned from the genome of Drosophila melanogaster, reveals a structural similarity with the proviral form of the retroviruses including a pol-like gene containing a putative reverse-transcriptase(RT)-coding sequence. Diverse parts of that sequence were subcloned and expressed in Escherichia coli. It has been demonstrated that the expression of the RT-like sequence, when translated, gives rise to peptides displaying enzyme activity characteristic of a true RT enzyme. In addition, rabbit antisera directed against such recombinant proteins allowed us to detect an immunoreactive protein of around 110 kDa, which was only present in D. melanogaster cell lines, but not in cells derived from Drosophila virilis or Drosophila hydei, whose genomes do not bear the 1731 element. This protein is expected to correspond to a non-processed pol-gene translated product and cosediments with virus-like particles exhibiting RT activity.
Champion
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Institut de Chimie Biologique, Université de Provence, Marseille, France.
Maisonhaute
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Kim
M H
MH
Best-Belpomme
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M
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification / Q000378:metabolism; D004330:Drosophila / Q000201:enzymology / Q000235:genetics; D004331:Drosophila melanogaster / Q000201:enzymology* / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D012194:RNA-Directed DNA Polymerase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D012190:Retroviridae / Q000201:enzymology / Q000235:genetics*
D004251:DNA Transposable Elements; D011994:Recombinant Proteins; D012194:RNA-Directed DNA Polymerase; D004259:DNA-Directed DNA Polymerase