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J. Biol. Chem., Vol. 277, Issue 20, 17877-17882, May 17, 2002
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,
From the Institut de Génétique Humaine, CNRS, 141 Rue
de la Cardonille, 34396 Montpellier Cedex 5, France
Non-long terminal repeat retrotransposons,
widespread among eukaryotic genomes, transpose by reverse transcription
of an RNA intermediate. Some of them, like L1 in the human, terminate
at the 3'-end with a poly(dA) stretch whereas 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. Here, we have used an RNA circularization/reverse transcription-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. These transcripts are capped and
polyadenylated similar to conventional messenger RNAs. We have analyzed
the 3'-ends of transcripts and transposed copies produced by I elements
mutated at the 3'-ends. Transcripts devoid of tandem UAA repeats,
although capable of building the components of the retrotransposition
machinery, are inefficiently used as retrotransposition intermediates.
Such transcripts produce rare new integrated copies issued from the
inaccurate 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 the 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.
Recipient of fellowships from the Fondation pour la Recherche
Médicale (FRM) and the Ministère de la Recherche et de la Technologie.
§
To whom correspondence should be addressed. Tel.: 4-99-61-99-48;
Fax: 4-99-61-99-01; E-mail: busseau@igh.cnrs.fr.
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