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. 1995 Dec;69(12):7845–7850. doi: 10.1128/jvi.69.12.7845-7850.1995

Plus-strand strong-stop DNA synthesis in retrotransposon Ty1.

V Lauermann 1, K Nam 1, J Trambley 1, J D Boeke 1
PMCID: PMC189728  PMID: 7494296

Abstract

Reverse transcription in the yeast retrotransposon Ty1 follows the general "rules" of retroviral replication overall. However, some details of the retroviral and Ty1 reverse transcription processes are different. We have identified and determined the structure of plus-strand strong-stop DNA and examined the effect of polypurine tract deletion mutations on its synthesis. Furthermore, we have defined the stop signal for plus-strand strong-stop DNA synthesis as an unusual 2'-O-ribosylated nucleotide in the primer tRNA. Full-length plus-strand strong-stop DNA, following strand transfer, would have a terminal 2-base mismatch with minus-strand DNA. These findings indicate that the mechanism of plus-strand strong-stop DNA transfer in Ty1 differs from that of the retroviral transfer and suggest that full-length plus-strand strong-stop DNA is not a direct intermediate in Ty1 retrotransposition.

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Selected References

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  1. Aström S. U., Byström A. S. Rit1, a tRNA backbone-modifying enzyme that mediates initiator and elongator tRNA discrimination. Cell. 1994 Nov 4;79(3):535–546. doi: 10.1016/0092-8674(94)90262-3. [DOI] [PubMed] [Google Scholar]
  2. Aström S. U., von Pawel-Rammingen U., Byström A. S. The yeast initiator tRNAMet can act as an elongator tRNA(Met) in vivo. J Mol Biol. 1993 Sep 5;233(1):43–58. doi: 10.1006/jmbi.1993.1483. [DOI] [PubMed] [Google Scholar]
  3. Basavappa R., Sigler P. B. The 3 A crystal structure of yeast initiator tRNA: functional implications in initiator/elongator discrimination. EMBO J. 1991 Oct;10(10):3105–3111. doi: 10.1002/j.1460-2075.1991.tb07864.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boeke J. D., Chapman K. B. Retrotransposition mechanisms. Curr Opin Cell Biol. 1991 Jun;3(3):502–507. doi: 10.1016/0955-0674(91)90079-e. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Boeke J. D., Styles C. A., Fink G. R. Saccharomyces cerevisiae SPT3 gene is required for transposition and transpositional recombination of chromosomal Ty elements. Mol Cell Biol. 1986 Nov;6(11):3575–3581. doi: 10.1128/mcb.6.11.3575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Braiterman L. T., Monokian G. M., Eichinger D. J., Merbs S. L., Gabriel A., Boeke J. D. In-frame linker insertion mutagenesis of yeast transposon Ty1: phenotypic analysis. Gene. 1994 Feb 11;139(1):19–26. doi: 10.1016/0378-1119(94)90518-5. [DOI] [PubMed] [Google Scholar]
  9. Chapman K. B., Boeke J. D. Isolation and characterization of the gene encoding yeast debranching enzyme. Cell. 1991 May 3;65(3):483–492. doi: 10.1016/0092-8674(91)90466-c. [DOI] [PubMed] [Google Scholar]
  10. Chapman K. B., Byström A. S., Boeke J. D. Initiator methionine tRNA is essential for Ty1 transposition in yeast. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3236–3240. doi: 10.1073/pnas.89.8.3236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Clare J., Farabaugh P. Nucleotide sequence of a yeast Ty element: evidence for an unusual mechanism of gene expression. Proc Natl Acad Sci U S A. 1985 May;82(9):2829–2833. doi: 10.1073/pnas.82.9.2829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Desgrès J., Keith G., Kuo K. C., Gehrke C. W. Presence of phosphorylated O-ribosyl-adenosine in T-psi-stem of yeast methionine initiator tRNA. Nucleic Acids Res. 1989 Feb 11;17(3):865–882. doi: 10.1093/nar/17.3.865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gilboa E., Mitra S. W., Goff S., Baltimore D. A detailed model of reverse transcription and tests of crucial aspects. Cell. 1979 Sep;18(1):93–100. doi: 10.1016/0092-8674(79)90357-x. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Inouye S., Hsu M. Y., Eagle S., Inouye M. Reverse transcriptase associated with the biosynthesis of the branched RNA-linked msDNA in Myxococcus xanthus. Cell. 1989 Feb 24;56(4):709–717. doi: 10.1016/0092-8674(89)90593-x. [DOI] [PubMed] [Google Scholar]
  16. Kiesewetter S., Ott G., Sprinzl M. The role of modified purine 64 in initiator/elongator discrimination of tRNA(iMet) from yeast and wheat germ. Nucleic Acids Res. 1990 Aug 25;18(16):4677–4682. doi: 10.1093/nar/18.16.4677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lampson B. C., Inouye M., Inouye S. Reverse transcriptase with concomitant ribonuclease H activity in the cell-free synthesis of branched RNA-linked msDNA of Myxococcus xanthus. Cell. 1989 Feb 24;56(4):701–707. doi: 10.1016/0092-8674(89)90592-8. [DOI] [PubMed] [Google Scholar]
  19. Lauermann V., Boeke J. D. The primer tRNA sequence is not inherited during Ty1 retrotransposition. Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):9847–9851. doi: 10.1073/pnas.91.21.9847. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Nam K., Hudson R. H., Chapman K. B., Ganeshan K., Damha M. J., Boeke J. D. Yeast lariat debranching enzyme. Substrate and sequence specificity. J Biol Chem. 1994 Aug 12;269(32):20613–20621. [PubMed] [Google Scholar]
  22. Natsoulis G., Thomas W., Roghmann M. C., Winston F., Boeke J. D. Ty1 transposition in Saccharomyces cerevisiae is nonrandom. Genetics. 1989 Oct;123(2):269–279. doi: 10.1093/genetics/123.2.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Rattray A. J., Champoux J. J. Plus-strand priming by Moloney murine leukemia virus. The sequence features important for cleavage by RNase H. J Mol Biol. 1989 Aug 5;208(3):445–456. doi: 10.1016/0022-2836(89)90508-1. [DOI] [PubMed] [Google Scholar]
  25. Shimada M., Hosaka H., Takaku H., Smith J. S., Roth M. J., Inouye S., Inouye M. Specificity of priming reaction of HIV-1 reverse transcriptase, 2'-OH or 3'-OH. J Biol Chem. 1994 Feb 11;269(6):3925–3927. [PubMed] [Google Scholar]
  26. Sprinzl M., Dank N., Nock S., Schön A. Compilation of tRNA sequences and sequences of tRNA genes. Nucleic Acids Res. 1991 Apr 25;19 (Suppl):2127–2171. doi: 10.1093/nar/19.suppl.2127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Taylor J. M., Hsu T. W., Yeater C., Mason W. S. Synthesis and integration of avian sarcoma virus DNA. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 2):1091–1096. doi: 10.1101/sqb.1980.044.01.117. [DOI] [PubMed] [Google Scholar]
  28. Winston F., Durbin K. J., Fink G. R. The SPT3 gene is required for normal transcription of Ty elements in S. cerevisiae. Cell. 1984 Dec;39(3 Pt 2):675–682. doi: 10.1016/0092-8674(84)90474-4. [DOI] [PubMed] [Google Scholar]
  29. Xu H., Boeke J. D. Localization of sequences required in cis for yeast Ty1 element transposition near the long terminal repeats: analysis of mini-Ty1 elements. Mol Cell Biol. 1990 Jun;10(6):2695–2702. doi: 10.1128/mcb.10.6.2695. [DOI] [PMC free article] [PubMed] [Google Scholar]

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