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M200996200v1
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Papers In Press, published online ahead of print March 6, 2002
J. Biol. Chem, 10.1074/jbc.M200996200
Submitted on January 30, 2002
Revised on March 5, 2002
Accepted on March 6, 2002

Tandem UAA repeats at the 3' end of the transcript are essential for precise initiation of reverse transcription of the I factor in Drosophila melanogaster

Séverine Chambeyron, Alain Bucheton, and Isabelle Busseau

IGH, CNRS, Montpellier 34396

Corresponding Author: busseau{at}igh.cnrs.fr

Non-LTR retrotransposons, widespread among eukaryotic genomes, transpose by reverse transcription of an RNA intermediate. Some of them, like L1 in human, terminate at the 3' end with a poly(dA) stretch, while others, like the I factor in Drosophila melanogaster, have instead a short sequence repeated in tandem. This suggests different requirements for the initiation of reverse transcription. We have used an RNA cicularization/RT-PCR technique to analyze the 5' and 3' ends of the full length transcripts produced by the I factor at the time of active retrotransposition. We find that these transcripts are capped and polyadenylated as conventional messenger RNAs. We have analysed the 3’ ends of transcripts and transposed copies produced by I elements mutated at the 3’ ends. We find that transcripts devoid of tandem UAA repeats, although capable of building the components of the retrotransposition machinery, are inefficiently used as retrotransposition intermediates. They produce rare new integrated copies issued from random initiation of reverse transcription near the 3' end of the element. The tandem UAA repeats at the 3' end of the transcripts of I are required for efficient and precise initiation of reverse transcription. This strong specificity of the I factor reverse transcriptase for its own transcript has implications for the impact of I factor retrotransposition on the host genome.


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