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LINE-mediated retrotransposition of marked Alu sequences

Nature Genetics volume 35, pages 41–48 (2003)Cite this article

Abstract

Alu elements are the most successful transposons in humans. They are 300-bp non-coding sequences transcribed by RNA polymerase III (Pol III) and are expected to retrotranspose with the aid of reverse transcriptases of cellular origin. We previously showed that human LINEs can generate cDNA copies of any mRNA transcript by means of a retroposition process involving reverse transcription and integration by the LINE-encoded endonuclease and reverse transcriptase. Here we show mobility of marked Alu sequences in human HeLa cells with the canonical features of a retrotransposition process, including splicing out of an autocatalytic intron introduced into the marked sequence, target site duplications of varying lengths and integrations into consensus A-rich sequences. We further show that the poly-A stretch at the Alu 3′ end is essential for mobility, that LINEs are required for transposition and that the rate of retroposition is 100–1,000 times higher for Alu transcripts than for control mRNAs, thus accounting for the high mutational activity of these elements observed in humans.

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Figure 1: Structure of the Alu SINE and rationale of the assay for retrotransposition.
Figure 2: High frequency L1-mediated Alu retrotransposition.
Figure 3: Structure of retrotransposed marked Alu sequences.
Figure 4: Chromosomal localization and properties of five de novo marked Alu insertions.
Figure 5: Elements required in trans for Alu retrotransposition.
Figure 6: Alu sequences required in cis for retrotransposition: role of the Alu poly-A tract and internal sequences.
Figure 7: Alu retrotransposition: cis and trans effects of the LINE-assisted processes.

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Acknowledgements

We thank H. Kazazian, M. Wallace and E. Ullu for the L1-RP, Alu-NF1, and 7SL RNA gene plasmids, respectively; J. Maestre and P. Dessen for discussions; and C. Lavialle for comments and critical reading of the manuscript. This work was supported by the Centre National de la Recherche Scientifique and a grant from the Ligue Nationale Contre le Cancer (Equipe “labellisée”).

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  1. Unité des Rétrovirus Endogènes et Eléments Rétroïdes des Eucaryotes Supérieurs, UMR 8122 CNRS, Institut Gustave Roussy, 39 rue Camille Desmoulins, Villejuif, 94805, Cedex, France

    Marie Dewannieux, Cécile Esnault & Thierry Heidmann

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Dewannieux, M., Esnault, C. & Heidmann, T. LINE-mediated retrotransposition of marked Alu sequences. Nat Genet 35, 41–48 (2003). https://doi.org/10.1038/ng1223

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