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. 1990 Dec;126(4):837–850. doi: 10.1093/genetics/126.4.837

Transfer RNA Genes Are Genomic Targets for De Novo Transposition of the Yeast Retrotransposon Ty3

D L Chalker 1, S B Sandmeyer 1
PMCID: PMC1204282  PMID: 1963869

Abstract

Insertions of the yeast element Ty3 resulting from induced retrotransposition were characterized in order to identify the genomic targets of transposition. The DNA sequences of the junctions between Ty3 and flanking DNA were determined for two insertions of an unmarked element. Each insertion was at position -17 from the 5' end of a tRNA-coding sequence. Ninety-one independent insertions of a marked Ty3 element were studied by Southern blot analysis. Pairs of independent insertions into seven genomic loci accounted for 14 of these insertions. The DNA sequence flanking the insertion site was determined for at least one member of each pair of integrated elements. In each case, insertion was at position -16 or -17 relative to the 5' end of one of seven different tRNA genes. This proportion of genomic loci used twice for Ty3 integration is consistent with that predicted by a Poisson distribution for a number of genomic targets roughly equivalent to the estimated number of yeast tRNA genes. In addition, insertions upstream of the same tRNA gene in one case were at different positions, but in all cases were in the same orientation. Thus, genomic insertions of Ty3 in a particular orientation are apparently specified by the target, while the actual position of the insertion relative to the tRNA-coding sequence can vary slightly.

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Selected References

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  1. Baker R. E., Eigel A., Vögtel D., Feldmann H. Nucleotide sequences of yeast genes for tRNA(2), tRNA(2) and tRNA(1): homology blocks occur in the vicinity of different tRNA genes. EMBO J. 1982;1(3):291–295. doi: 10.1002/j.1460-2075.1982.tb01162.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bell G. I., DeGennaro L. J., Gelfand D. H., Bishop R. J., Valenzuela P., Rutter W. J. Ribosomal RNA genes of Saccharomyces cerevisiae. I. Physical map of the repeating unit and location of the regions coding for 5 S, 5.8 S, 18 S, and 25 S ribosomal RNAs. J Biol Chem. 1977 Nov 25;252(22):8118–8125. [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. Boeke J. D., LaCroute F., Fink G. R. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet. 1984;197(2):345–346. doi: 10.1007/BF00330984. [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. Braun B. R., Riggs D. L., Kassavetis G. A., Geiduschek E. P. Multiple states of protein-DNA interaction in the assembly of transcription complexes on Saccharomyces cerevisiae 5S ribosomal RNA genes. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2530–2534. doi: 10.1073/pnas.86.8.2530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Brodeur G. M., Sandmeyer S. B., Olson M. V. Consistent association between sigma elements and tRNA genes in yeast. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3292–3296. doi: 10.1073/pnas.80.11.3292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Brow D. A., Guthrie C. Spliceosomal RNA U6 is remarkably conserved from yeast to mammals. Nature. 1988 Jul 21;334(6179):213–218. doi: 10.1038/334213a0. [DOI] [PubMed] [Google Scholar]
  11. Brown P. O., Bowerman B., Varmus H. E., Bishop J. M. Correct integration of retroviral DNA in vitro. Cell. 1987 May 8;49(3):347–356. doi: 10.1016/0092-8674(87)90287-x. [DOI] [PubMed] [Google Scholar]
  12. Brown P. O., Bowerman B., Varmus H. E., Bishop J. M. Retroviral integration: structure of the initial covalent product and its precursor, and a role for the viral IN protein. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2525–2529. doi: 10.1073/pnas.86.8.2525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Donehower L. A. Analysis of mutant Moloney murine leukemia viruses containing linker insertion mutations in the 3' region of pol. J Virol. 1988 Nov;62(11):3958–3964. doi: 10.1128/jvi.62.11.3958-3964.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Donehower L. A., Varmus H. E. A mutant murine leukemia virus with a single missense codon in pol is defective in a function affecting integration. Proc Natl Acad Sci U S A. 1984 Oct;81(20):6461–6465. doi: 10.1073/pnas.81.20.6461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Eigel A., Feldmann H. Ty1 and delta elements occur adjacent to several tRNA genes in yeast. EMBO J. 1982;1(10):1245–1250. doi: 10.1002/j.1460-2075.1982.tb00020.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  18. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  19. Goodman H. M., Olson M. V., Hall B. D. Nucleotide sequence of a mutant eukaryotic gene: the yeast tyrosine-inserting ochre suppressor SUP4-o. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5453–5457. doi: 10.1073/pnas.74.12.5453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Haltiner M., Kempe T., Tjian R. A novel strategy for constructing clustered point mutations. Nucleic Acids Res. 1985 Feb 11;13(3):1015–1025. doi: 10.1093/nar/13.3.1015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Hansen L. J., Sandmeyer S. B. Characterization of a transpositionally active Ty3 element and identification of the Ty3 integrase protein. J Virol. 1990 Jun;64(6):2599–2607. doi: 10.1128/jvi.64.6.2599-2607.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hauber J., Stucka R., Krieg R., Feldmann H. Analysis of yeast chromosomal regions carrying members of the glutamate tRNA gene family: various transposable elements are associated with them. Nucleic Acids Res. 1988 Nov 25;16(22):10623–10634. doi: 10.1093/nar/16.22.10623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ikenaga H., Saigo K. Insertion of a movable genetic element, 297, into the T-A-T-A box for the H3 histone gene in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4143–4147. doi: 10.1073/pnas.79.13.4143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Inouye S., Yuki S., Saigo K. Sequence-specific insertion of the Drosophila transposable genetic element 17.6. 1984 Jul 26-Aug 1Nature. 310(5975):332–333. doi: 10.1038/310332a0. [DOI] [PubMed] [Google Scholar]
  26. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Joyce C. M., Grindley N. D. Method for determining whether a gene of Escherichia coli is essential: application to the polA gene. J Bacteriol. 1984 May;158(2):636–643. doi: 10.1128/jb.158.2.636-643.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Krol A., Carbon P., Ebel J. P., Appel B. Xenopus tropicalis U6 snRNA genes transcribed by Pol III contain the upstream promoter elements used by Pol II dependent U snRNA genes. Nucleic Acids Res. 1987 Mar 25;15(6):2463–2478. doi: 10.1093/nar/15.6.2463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kunkel G. R., Maser R. L., Calvet J. P., Pederson T. U6 small nuclear RNA is transcribed by RNA polymerase III. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8575–8579. doi: 10.1073/pnas.83.22.8575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Marschalek R., Brechner T., Amon-Böhm E., Dingermann T. Transfer RNA genes: landmarks for integration of mobile genetic elements in Dictyostelium discoideum. Science. 1989 Jun 23;244(4911):1493–1496. doi: 10.1126/science.2567533. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Nelböck P., Stucka R., Feldmann H. Different patterns of transposable elements in the vicinity of tRNA genes in yeast: a possible clue to transcriptional modulation. Biol Chem Hoppe Seyler. 1985 Nov;366(11):1041–1051. doi: 10.1515/bchm3.1985.366.2.1041. [DOI] [PubMed] [Google Scholar]
  34. Olah J., Feldmann H. Structure of a yeast non-initiating methionine-tRNA gene. Nucleic Acids Res. 1980 May 10;8(9):1975–1986. doi: 10.1093/nar/8.9.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Petes T. D. Yeast ribosomal DNA genes are located on chromosome XII. Proc Natl Acad Sci U S A. 1979 Jan;76(1):410–414. doi: 10.1073/pnas.76.1.410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Reddy R., Henning D., Das G., Harless M., Wright D. The capped U6 small nuclear RNA is transcribed by RNA polymerase III. J Biol Chem. 1987 Jan 5;262(1):75–81. [PubMed] [Google Scholar]
  37. Reddy R. Transcription of a U6 small nuclear RNA gene in vitro. Transcription of a mouse U6 small nuclear RNA gene in vitro by RNA polymerase III is dependent on transcription factor(s) different from transcription factors IIIA, IIIB, and IIIC. J Biol Chem. 1988 Nov 5;263(31):15980–15984. [PubMed] [Google Scholar]
  38. Robinson H. L., Gagnon G. C. Patterns of proviral insertion and deletion in avian leukosis virus-induced lymphomas. J Virol. 1986 Jan;57(1):28–36. doi: 10.1128/jvi.57.1.28-36.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Rohdewohld H., Weiher H., Reik W., Jaenisch R., Breindl M. Retrovirus integration and chromatin structure: Moloney murine leukemia proviral integration sites map near DNase I-hypersensitive sites. J Virol. 1987 Feb;61(2):336–343. doi: 10.1128/jvi.61.2.336-343.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Roth M. J., Schwartzberg P. L., Goff S. P. Structure of the termini of DNA intermediates in the integration of retroviral DNA: dependence on IN function and terminal DNA sequence. Cell. 1989 Jul 14;58(1):47–54. doi: 10.1016/0092-8674(89)90401-7. [DOI] [PubMed] [Google Scholar]
  41. Sandmeyer S. B., Bilanchone V. W., Clark D. J., Morcos P., Carle G. F., Brodeur G. M. Sigma elements are position-specific for many different yeast tRNA genes. Nucleic Acids Res. 1988 Feb 25;16(4):1499–1515. doi: 10.1093/nar/16.4.1499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sandmeyer S. B., Olson M. V. Insertion of a repetitive element at the same position in the 5'-flanking regions of two dissimilar yeast tRNA genes. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7674–7678. doi: 10.1073/pnas.79.24.7674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Schwartzberg P., Colicelli J., Goff S. P. Construction and analysis of deletion mutations in the pol gene of Moloney murine leukemia virus: a new viral function required for productive infection. Cell. 1984 Jul;37(3):1043–1052. doi: 10.1016/0092-8674(84)90439-2. [DOI] [PubMed] [Google Scholar]
  45. Schweizer E., MacKechnie C., Halvorson H. O. The redundancy of ribosomal and transfer RNA genes in Saccharomyces cerevisiae. J Mol Biol. 1969 Mar 14;40(2):261–277. doi: 10.1016/0022-2836(69)90474-4. [DOI] [PubMed] [Google Scholar]
  46. Shih C. C., Stoye J. P., Coffin J. M. Highly preferred targets for retrovirus integration. Cell. 1988 May 20;53(4):531–537. doi: 10.1016/0092-8674(88)90569-7. [DOI] [PubMed] [Google Scholar]
  47. Struhl K., Stinchcomb D. T., Scherer S., Davis R. W. High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1035–1039. doi: 10.1073/pnas.76.3.1035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Stucka R., Lochmüller H., Feldmann H. Ty4, a novel low-copy number element in Saccharomyces cerevisiae: one copy is located in a cluster of Ty elements and tRNA genes. Nucleic Acids Res. 1989 Jul 11;17(13):4993–5001. doi: 10.1093/nar/17.13.4993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Tanda S., Shrimpton A. E., Chueh L. L., Itayama H., Matsubayashi H., Saigo K., Tobari Y. N., Langley C. H. Retrovirus-like features and site specific insertions of a transposable element, tom, in Drosophila ananassae. Mol Gen Genet. 1988 Nov;214(3):405–411. doi: 10.1007/BF00330473. [DOI] [PubMed] [Google Scholar]
  50. Tschumper G., Carbon J. Delta sequences and double symmetry in a yeast chromosomal replicator region. J Mol Biol. 1982 Apr 5;156(2):293–307. doi: 10.1016/0022-2836(82)90330-8. [DOI] [PubMed] [Google Scholar]
  51. Van Arsdell S. W., Stetler G. L., Thorner J. The yeast repeated element sigma contains a hormone-inducible promoter. Mol Cell Biol. 1987 Feb;7(2):749–759. doi: 10.1128/mcb.7.2.749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Vijaya S., Steffen D. L., Robinson H. L. Acceptor sites for retroviral integrations map near DNase I-hypersensitive sites in chromatin. J Virol. 1986 Nov;60(2):683–692. doi: 10.1128/jvi.60.2.683-692.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Weissenbach J., Martin R., Dirheimer G. The primary structure of tRNAIIAgr from brewers' yeast. 2. Partial digestion with ribonuclease T1 and derivation of the complete sequence. Eur J Biochem. 1975 Aug 15;56(2):527–532. doi: 10.1111/j.1432-1033.1975.tb02258.x. [DOI] [PubMed] [Google Scholar]
  54. Wilke C. M., Heidler S. H., Brown N., Liebman S. W. Analysis of yeast retrotransposon Ty insertions at the CAN1 locus. Genetics. 1989 Dec;123(4):655–665. doi: 10.1093/genetics/123.4.655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. del Rey F. J., Donahue T. F., Fink G. R. sigma, a repetitive element found adjacent to tRNA genes of yeast. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4138–4142. doi: 10.1073/pnas.79.13.4138. [DOI] [PMC free article] [PubMed] [Google Scholar]

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