Search tips
Search criteria 


Logo of jvirolPermissionsJournals.ASM.orgJournalJV ArticleJournal InfoAuthorsReviewers
J Virol. 1995 December; 69(12): 7845–7850.
PMCID: PMC189728

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


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.

Full Text

The Full Text of this article is available as a PDF (352K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Aström SU, von Pawel-Rammingen U, Byström AS. The yeast initiator tRNAMet can act as an elongator tRNA(Met) in vivo. J Mol Biol. 1993 Sep 5;233(1):43–58. [PubMed]
  • Aström SU, Byström AS. Rit1, a tRNA backbone-modifying enzyme that mediates initiator and elongator tRNA discrimination. Cell. 1994 Nov 4;79(3):535–546. [PubMed]
  • Basavappa R, Sigler PB. The 3 A crystal structure of yeast initiator tRNA: functional implications in initiator/elongator discrimination. EMBO J. 1991 Oct;10(10):3105–3111. [PubMed]
  • Boeke JD, Chapman KB. Retrotransposition mechanisms. Curr Opin Cell Biol. 1991 Jun;3(3):502–507. [PubMed]
  • Boeke JD, Garfinkel DJ, Styles CA, Fink GR. Ty elements transpose through an RNA intermediate. Cell. 1985 Mar;40(3):491–500. [PubMed]
  • Boeke JD, Styles CA, Fink GR. Saccharomyces cerevisiae SPT3 gene is required for transposition and transpositional recombination of chromosomal Ty elements. Mol Cell Biol. 1986 Nov;6(11):3575–3581. [PMC free article] [PubMed]
  • Boeke JD, Xu H, Fink GR. A general method for the chromosomal amplification of genes in yeast. Science. 1988 Jan 15;239(4837):280–282. [PubMed]
  • Braiterman LT, Monokian GM, Eichinger DJ, Merbs SL, Gabriel A, Boeke JD. In-frame linker insertion mutagenesis of yeast transposon Ty1: phenotypic analysis. Gene. 1994 Feb 11;139(1):19–26. [PubMed]
  • Chapman KB, Boeke JD. Isolation and characterization of the gene encoding yeast debranching enzyme. Cell. 1991 May 3;65(3):483–492. [PubMed]
  • Chapman KB, Byström AS, Boeke JD. Initiator methionine tRNA is essential for Ty1 transposition in yeast. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3236–3240. [PubMed]
  • 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. [PubMed]
  • Desgrès J, Keith G, Kuo KC, Gehrke CW. 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. [PMC free article] [PubMed]
  • Gilboa E, Mitra SW, Goff S, Baltimore D. A detailed model of reverse transcription and tests of crucial aspects. Cell. 1979 Sep;18(1):93–100. [PubMed]
  • Hansen LJ, Chalker DL, Sandmeyer SB. Ty3, a yeast retrotransposon associated with tRNA genes, has homology to animal retroviruses. Mol Cell Biol. 1988 Dec;8(12):5245–5256. [PMC free article] [PubMed]
  • Inouye S, Hsu MY, 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. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Kunkel TA. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. [PubMed]
  • Lampson BC, 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. [PubMed]
  • Lauermann V, Boeke JD. The primer tRNA sequence is not inherited during Ty1 retrotransposition. Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):9847–9851. [PubMed]
  • 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. [PubMed]
  • Nam K, Hudson RH, Chapman KB, Ganeshan K, Damha MJ, Boeke JD. Yeast lariat debranching enzyme. Substrate and sequence specificity. J Biol Chem. 1994 Aug 12;269(32):20613–20621. [PubMed]
  • Natsoulis G, Thomas W, Roghmann MC, Winston F, Boeke JD. Ty1 transposition in Saccharomyces cerevisiae is nonrandom. Genetics. 1989 Oct;123(2):269–279. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Rattray AJ, Champoux JJ. 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. [PubMed]
  • Shimada M, Hosaka H, Takaku H, Smith JS, Roth MJ, 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]
  • 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. [PMC free article] [PubMed]
  • Taylor JM, Hsu TW, Yeater C, Mason WS. Synthesis and integration of avian sarcoma virus DNA. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 2):1091–1096. [PubMed]
  • Winston F, Durbin KJ, Fink GR. The SPT3 gene is required for normal transcription of Ty elements in S. cerevisiae. Cell. 1984 Dec;39(3 Pt 2):675–682. [PubMed]
  • Xu H, Boeke JD. 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. [PMC free article] [PubMed]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)