Heterogeneous terminal structure of Ty1 and Ty3 reverse transcripts

A specific terminal structure of preintegrative DNA is required for transposition of retroviruses and LTR-retrotransposons. We have used an anchored PCR technique to map the 3′ ends of DNA intermediates synthesized inside yeast Ty1 and Ty3 retrotransposon virus-like particles. We find that, unlike r...

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Bibliographic Details
Published in:Nucleic acids research 1997-06, Vol.25 (11), p.2161-2166
Main Authors: Wilhelm, M, Heyman, T, Friant, S, Wilhelm, F.X
Format: Article
Language:English
Subjects:
Cellular Biology
DNA
DNA Replication
DNA, Fungal
DNA, Fungal - chemistry
DNA, Fungal - metabolism
Life Sciences
Nucleic Acid Conformation
Nucleic Acid Hybridization
nucleotide sequences
Plasmids
Plasmids - chemistry
Plasmids - genetics
Plasmids - metabolism
Polymerase Chain Reaction
Retroelements
Retroelements - genetics
retrotransposons
reverse transcription
Ribonuclease H - metabolism
Ribonuclease H, Calf Thymus
RNA, Transfer
RNA, Transfer - chemistry
Saccharomyces cerevisiae
Transcription, Genetic
virus-like particles
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Summary:A specific terminal structure of preintegrative DNA is required for transposition of retroviruses and LTR-retrotransposons. We have used an anchored PCR technique to map the 3′ ends of DNA intermediates synthesized inside yeast Ty1 and Ty3 retrotransposon virus-like particles. We find that, unlike retroviruses, Ty1 replicated DNA does not have two extra base pairs at its 3′ ends. In contrast some Ty3 preintegrative DNA molecules have two extra nucleotides at the 3′ end of upstream and downstream long terminal repeats. Moreover we find that some molecules of replicated Ty3 DNA have more than two extra nucleotides at the 3′ end of the upstream LTR. This observation could be accounted for by imprecise RNAse H cutting of the PPT sequence. The site of Ty1 and Ty3 plus-strand strongstop DNA termination was also examined. Our results confirm that the prominent Ty1 and Ty3 plus-strand strong-stop molecules harbor 12 tRNA templated bases but also show that some Ty1 and Ty3 plusstrand strong-stop DNA molecules harbor less tRNA templated bases. We propose that these less than full length plus-strand molecules could be active intermediates in Ty retrotransposon replication.
ISSN:0305-1048
1362-4962
1362-4962
DOI:10.1093/nar/25.11.2161