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Published online 23 January 2004

Nucleic Acids Research, 2004, Vol. 32, No. 2 502-510
© 2004 Oxford University Press

More active human L1 retrotransposons produce longer insertions

Alexander H. Farley, Eline T. Luning Prak1 and Haig H. Kazazian Jr*

Department of Genetics, and 1 Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, PA, USA

*To whom correspondence should be addressed. Tel: +1 215 898 3582; Fax: +1 215 573 7760; Email: kazazian{at}mail.med.upenn.edu
Correspondence may also be addressed to Eline T. Luning Prak. Tel: +1 215 746 5768; Fax: +1 215 573 6317; Email: luning{at}mail.med.upenn.edu

The vast majority of L1 insertions are 5' truncated and thus inactive. Yet, the mechanism of 5' truncation is unknown. To examine whether the frequency of L1 retrotransposition is directly correlated with the length of genomic L1 insertions, we used a cell culture assay to measure retrotransposition frequency and a PCR-based assay to measure L1 insertion length. We tested five full-length human L1 elements that retrotranspose at different frequencies: LRE3, L1RP, L1.3, L1.2A and L1.2B. Our data suggest that L1 insertion length correlates with L1 retrotransposition frequency for insertions >1 kb in length. For two elements, L1RP and L1.2A, we found that swapping the reverse transcriptase domains had little effect. Instead, we found that genomic insertion length and retrotransposition frequency are substantially affected by amino acid substitutions at positions 363, 1220 and 1259 in ORF2. We suggest that the region containing residues 1220 and 1259 may be important in the binding of ORF2p to L1 RNA to facilitate reverse transcription.


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