Authentic reverse transcriptase is coded by jockey, a mobile Drosophila element related to mammalian LINEs.
The mobile element jockey is similar in structural organization and coding potential to the LINEs of various organisms. It is transcribed at different stages of Drosophila ontogenesis. The Drosophila LINE family includes active transposable elements. Current models for the mechanism of transposition involve reverse transcription of an RNA intermediate and utilization of element-encoded proteins. As demonstrated here, a 2.23 kb DNA fragment from the region of jockey encoding the putative reverse transcriptase was stably introduced into an expression system under inducible control of the Escherichia coli lac regulatory elements. We describe the expression of the 92 kDa protein and identify this polypeptide alone as the authentic jockey reverse transcriptase based on some of its physical and enzymic properties. The jockey polymerase demonstrates RNA and DNA-directed DNA polymerase activities but lacks detectable RNase H, has a temperature optimum at 26 degrees C, requires Mg2+ or Mn2+ as a cofactor and is inactivated by sulphydryl reagent. The enzyme prefers poly(rC) and poly(rA) as template and 'activated' DNA is not effective.
Ivanov
V A
VA
Institute of Cell Biophysics, Academy of Sciences, Poustchino, Moscow Region, USSR.
Melnikov
A A
AA
Siunov
A V
AV
Fodor
I I
II
Ilyin
Y V
YV
D000818:Animals; D002848:Chromatography, DEAE-Cellulose; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004251:DNA Transposable Elements; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004331:Drosophila melanogaster; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004722:Endoribonucleases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D010957:Plasmids; D012194:RNA-Directed DNA Polymerase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D016914:Ribonuclease H; D013696:Temperature; D013698:Templates, Genetic
D004251:DNA Transposable Elements; D004247:DNA; D012194:RNA-Directed DNA Polymerase; D012321:DNA-Directed RNA Polymerases; D004722:Endoribonucleases; D016914:Ribonuclease H