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. 1997 Jun 1;25(11):2161–2166. doi: 10.1093/nar/25.11.2161

Heterogeneous terminal structure of Ty1 and Ty3 reverse transcripts.

M Wilhelm 1, T Heyman 1, S Friant 1, F X Wilhelm 1
PMCID: PMC146723  PMID: 9153316

Abstract

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 strong-stop 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 plus-strand 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.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Boeke J. D., Eichinger D., Castrillon D., Fink G. R. The Saccharomyces cerevisiae genome contains functional and nonfunctional copies of transposon Ty1. Mol Cell Biol. 1988 Apr;8(4):1432–1442. doi: 10.1128/mcb.8.4.1432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boeke J. D., Garfinkel D. J., Styles C. A., Fink G. R. Ty elements transpose through an RNA intermediate. Cell. 1985 Mar;40(3):491–500. doi: 10.1016/0092-8674(85)90197-7. [DOI] [PubMed] [Google Scholar]
  3. Boeke J. D., Xu H., Fink G. R. A general method for the chromosomal amplification of genes in yeast. Science. 1988 Jan 15;239(4837):280–282. doi: 10.1126/science.2827308. [DOI] [PubMed] [Google Scholar]
  4. Charneau P., Mirambeau G., Roux P., Paulous S., Buc H., Clavel F. HIV-1 reverse transcription. A termination step at the center of the genome. J Mol Biol. 1994 Sep 2;241(5):651–662. doi: 10.1006/jmbi.1994.1542. [DOI] [PubMed] [Google Scholar]
  5. Eichinger D. J., Boeke J. D. The DNA intermediate in yeast Ty1 element transposition copurifies with virus-like particles: cell-free Ty1 transposition. Cell. 1988 Sep 23;54(7):955–966. doi: 10.1016/0092-8674(88)90110-9. [DOI] [PubMed] [Google Scholar]
  6. Gabriel A., Willems M., Mules E. H., Boeke J. D. Replication infidelity during a single cycle of Ty1 retrotransposition. Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):7767–7771. doi: 10.1073/pnas.93.15.7767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hansen L. J., Chalker D. L., Sandmeyer S. B. Ty3, a yeast retrotransposon associated with tRNA genes, has homology to animal retroviruses. Mol Cell Biol. 1988 Dec;8(12):5245–5256. doi: 10.1128/mcb.8.12.5245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Heyman T., Agoutin B., Friant S., Wilhelm F. X., Wilhelm M. L. Plus-strand DNA synthesis of the yeast retrotransposon Ty1 is initiated at two sites, PPT1 next to the 3' LTR and PPT2 within the pol gene. PPT1 is sufficient for Ty1 transposition. J Mol Biol. 1995 Oct 20;253(2):291–303. doi: 10.1006/jmbi.1995.0553. [DOI] [PubMed] [Google Scholar]
  9. Katz R. A., Skalka A. M. The retroviral enzymes. Annu Rev Biochem. 1994;63:133–173. doi: 10.1146/annurev.bi.63.070194.001025. [DOI] [PubMed] [Google Scholar]
  10. Lauermann V., Nam K., Trambley J., Boeke J. D. Plus-strand strong-stop DNA synthesis in retrotransposon Ty1. J Virol. 1995 Dec;69(12):7845–7850. doi: 10.1128/jvi.69.12.7845-7850.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Müller F., Laufer W., Pott U., Ciriacy M. Characterization of products of TY1-mediated reverse transcription in Saccharomyces cerevisiae. Mol Gen Genet. 1991 Apr;226(1-2):145–153. doi: 10.1007/BF00273598. [DOI] [PubMed] [Google Scholar]
  12. Orlinsky K. J., Gu J., Hoyt M., Sandmeyer S., Menees T. M. Mutations in the Ty3 major homology region affect multiple steps in Ty3 retrotransposition. J Virol. 1996 Jun;70(6):3440–3448. doi: 10.1128/jvi.70.6.3440-3448.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Pochart P., Agoutin B., Rousset S., Chanet R., Doroszkiewicz V., Heyman T. Biochemical and electron microscope analyses of the DNA reverse transcripts present in the virus-like particles of the yeast transposon Ty1. Identification of a second origin of Ty1DNA plus strand synthesis. Nucleic Acids Res. 1993 Jul 25;21(15):3513–3520. doi: 10.1093/nar/21.15.3513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Varmus H. Retroviruses. Science. 1988 Jun 10;240(4858):1427–1435. doi: 10.1126/science.3287617. [DOI] [PubMed] [Google Scholar]

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