Analysis of the 5' junctions of R2 insertions with the 28S gene: implications for non-LTR retrotransposition.
R2 elements are non-long terminal repeat retrotransposable elements that insert into 28S rRNA genes of most insect species. The single open reading frame of R2 encodes a protein with both endonuclease activity, which cleaves the target site, and reverse transcriptase activity, which uses this cleavage to prime reverse transcription. This target-primed reverse transcription mechanism is also used by group II introns. Little is known of the mechanism by which the 5' end of R2 is integrated after reverse transcription. We have determined the 5' junction sequence of 94 R2 elements from 14 different species of Drosophila. Only 37% of the full-length elements contained precise 5' junctions; the remainder contained deletions of the 28S gene and/or insertions of additional sequences. Because the 5' junctions of truncated copies were similar to full-length elements, no sequences at the 5' end of R2 appear to be required for element integration. A model in which the R2 reverse transcriptase is capable of switching templates from the R2 RNA transcript to the upstream 28S gene can best explain the observed 5' junction sequences. This template jumping is analogous to the template switching of retroviral reverse transcriptases during formation of the double-stranded integration products.
George
J A
JA
Department of Biology, University of Rochester, New York 14627, USA.
Burke
W D
WD
Eickbush
T H
TH
D000818:Animals; D001483:Base Sequence; D004251:DNA Transposable Elements; D004275:DNA, Ribosomal; D004330:Drosophila / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics*; D017344:Genes, Insect; D008969:Molecular Sequence Data; D012339:RNA, Ribosomal, 28S; D012689:Sequence Homology, Nucleic Acid
D004251:DNA Transposable Elements; D004275:DNA, Ribosomal; D012339:RNA, Ribosomal, 28S