Genetics, Vol 147, 545-556, Copyright © 1997

INVESTIGATIONS

High Frequency cDNA Recombination of the Saccharomyces Retrotransposon Ty5: The LTR Mediates Formation of Tandem Elements

N. Ke and D. F. Voytas
Department of Zoology and Genetics, Iowa State University, Ames, Iowa 50011

Retroelement cDNA can integrate into the genome using the element-encoded integrase, or it can recombine with preexisting elements using the recombination system of the host. Recombination is a particularly important pathway for the yeast retrotransposon Ty5 and accounts for ~30% of the putative transposition events when a homologous substrate is carried on a plasmid and ~7% when the substrate is located at the chromosomal URA3 locus. Characterization of recombinants revealed that they are either simple replacements of the marker gene or tandem elements. Using an assay system in which the donor element and recombination substrates are separated, we found that the long terminal repeats (LTRs) are critical for tandem element formation. LTR-containing substrates generate tandem elements at frequencies more than 10-fold higher than similarly sized internal Ty5 sequences. Internal sequences, however, facilitate tandem element formation when associated with an LTR, and there is a linear relationship between frequencies of tandem element formation and the length of LTR-containing substrates. We propose that recombination is initiated between the LTRs of the cDNA and substrate and that internal sequences promote tandem element formation by facilitating sequence alignment. Because of its location in subtelomeric regions, recombinational amplification of Ty5 may contribute to the organization of chromosome ends.

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