Trans-complementation of an endonuclease-defective tagged I element as a tool for the study of retrotransposition in Drosophila melanogaster

• Published in: Molecular genetics and genomics : MGG Vol. 267; no. 6; pp. 829 - 834
• Main Authors: Robin, S, Chambeyron, S, Brun, C, Bucheton, A, Busseau, I
• Format: Journal Article
• Language: English
• Published: Germany Springer Nature B.V 01.08.2002; Springer Verlag
• Subjects: Animals; Animals, Genetically Modified; Chromosomes; Cloning; Crosses, Genetic; Drosophila melanogaster - genetics; Endonucleases - metabolism; Female; Genetic Complementation Test; Genetics; Genomes; Insects; Life Sciences; Male; Retroelements; Sequence Analysis, DNA
• ISSN: 1617-4615; 1617-4623
• DOI: 10.1007/s00438-002-0716-z

I factors are non-LTR retrotransposons of Drosophila melanogaster that transpose at high frequency in the germline of females resulting from appropriate crosses, allowing in vivo studies of the retrotransposition process. Reverse transcription of a full-length RNA intermediate is thought to occur at the site of integration, using a 3' hydroxyl group generated by endonucleolytic cleavage of the genomic DNA to prime synthesis of the first cDNA strand. This target-primed reverse transcription (TPRT) process is mediated by endonuclease and reverse transcriptase activities encoded by the element. We have designed a molecularly tagged, endonuclease-defective I element that can be mobilised with high efficiency by constructs that express the product of the I factor ORF2 in trans. This indicates that the endonuclease activity required for retrotransposition of the I factor can be provided in trans. Using this system, we show that the endonuclease domain of the R1Bm retrotransposon from Bombyx mori cannot functionally replace that of the I factor.