Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1995 May 15;14(10):2364–2373. doi: 10.1002/j.1460-2075.1995.tb07231.x

RNA-mediated transposition of the tobacco retrotransposon Tnt1 in Arabidopsis thaliana.

H Lucas 1, F Feuerbach 1, K Kunert 1, M A Grandbastien 1, M Caboche 1
PMCID: PMC398345  PMID: 7774594

Abstract

The tobacco (Nicotiana tabacum) retrotransposon Tnt1 was introduced into Arabidopsis thaliana. In this heterologous host plant species, Tnt1 undergoes an RNA-mediated transposition and creates a 5 bp duplication at the insertion sites. This is the first report of transposition of a retrotransposon after introduction into a heterologous host species. Tnt1 transposed during in vitro regeneration of transformed A.thaliana, but no transposition event was detected as happening in T2 and T3 generation plants. Newly synthesized copies of Tnt1 can integrate into coding regions of the host DNA. Our results open up the possibility of using Tnt1 as a new tool for insertional mutagenesis and functional analysis of plant genomes, in addition to the strategies of T-DNA and transposon tagging.

Full text

PDF
2364

Images in this article

2365Image
on p.2365
2366Image
on p.2366
2368Image
on p.2368
2368Image
on p.2368
2369Image
on p.2369

Selected References

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

  1. Aarts M. G., Dirkse W. G., Stiekema W. J., Pereira A. Transposon tagging of a male sterility gene in Arabidopsis. Nature. 1993 Jun 24;363(6431):715–717. doi: 10.1038/363715a0. [DOI] [PubMed] [Google Scholar]
  2. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  3. Ballinger D. G., Benzer S. Targeted gene mutations in Drosophila. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9402–9406. doi: 10.1073/pnas.86.23.9402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bancroft I., Dean C. Transposition pattern of the maize element Ds in Arabidopsis thaliana. Genetics. 1993 Aug;134(4):1221–1229. doi: 10.1093/genetics/134.4.1221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bancroft I., Jones J. D., Dean C. Heterologous transposon tagging of the DRL1 locus in Arabidopsis. Plant Cell. 1993 Jun;5(6):631–638. doi: 10.1105/tpc.5.6.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bevan M. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984 Nov 26;12(22):8711–8721. doi: 10.1093/nar/12.22.8711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bingham P. M., Levis R., Rubin G. M. Cloning of DNA sequences from the white locus of D. melanogaster by a novel and general method. Cell. 1981 Sep;25(3):693–704. doi: 10.1016/0092-8674(81)90176-8. [DOI] [PubMed] [Google Scholar]
  8. Brierley C., Flavell A. J. The retrotransposon copia controls the relative levels of its gene products post-transcriptionally by differential expression from its two major mRNAs. Nucleic Acids Res. 1990 May 25;18(10):2947–2951. doi: 10.1093/nar/18.10.2947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Camirand A., Brisson N. The complete nucleotide sequence of the Tst1 retrotransposon of potato. Nucleic Acids Res. 1990 Aug 25;18(16):4929–4929. doi: 10.1093/nar/18.16.4929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cardon G. H., Frey M., Saedler H., Gierl A. Definition and characterization of an artificial En/Spm-based transposon tagging system in transgenic tobacco. Plant Mol Biol. 1993 Oct;23(1):157–178. doi: 10.1007/BF00021428. [DOI] [PubMed] [Google Scholar]
  11. Casacuberta J. M., Grandbastien M. A. Characterisation of LTR sequences involved in the protoplast specific expression of the tobacco Tnt1 retrotransposon. Nucleic Acids Res. 1993 May 11;21(9):2087–2093. doi: 10.1093/nar/21.9.2087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Chalker D. L., Sandmeyer S. B. Ty3 integrates within the region of RNA polymerase III transcription initiation. Genes Dev. 1992 Jan;6(1):117–128. doi: 10.1101/gad.6.1.117. [DOI] [PubMed] [Google Scholar]
  13. Chandler V. L., Hardeman K. J. The Mu elements of Zea mays. Adv Genet. 1992;30:77–122. doi: 10.1016/s0065-2660(08)60319-3. [DOI] [PubMed] [Google Scholar]
  14. Chuck G., Robbins T., Nijjar C., Ralston E., Courtney-Gutterson N., Dooner H. K. Tagging and Cloning of a Petunia Flower Color Gene with the Maize Transposable Element Activator. Plant Cell. 1993 Apr;5(4):371–378. doi: 10.1105/tpc.5.4.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Curcio M. J., Hedge A. M., Boeke J. D., Garfinkel D. J. Ty RNA levels determine the spectrum of retrotransposition events that activate gene expression in Saccharomyces cerevisiae. Mol Gen Genet. 1990 Jan;220(2):213–221. doi: 10.1007/BF00260484. [DOI] [PubMed] [Google Scholar]
  16. Deblaere R., Bytebier B., De Greve H., Deboeck F., Schell J., Van Montagu M., Leemans J. Efficient octopine Ti plasmid-derived vectors for Agrobacterium-mediated gene transfer to plants. Nucleic Acids Res. 1985 Jul 11;13(13):4777–4788. doi: 10.1093/nar/13.13.4777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Denecke J., Botterman J., Deblaere R. Protein secretion in plant cells can occur via a default pathway. Plant Cell. 1990 Jan;2(1):51–59. doi: 10.1105/tpc.2.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Doolittle R. F., Feng D. F., Johnson M. S., McClure M. A. Origins and evolutionary relationships of retroviruses. Q Rev Biol. 1989 Mar;64(1):1–30. doi: 10.1086/416128. [DOI] [PubMed] [Google Scholar]
  20. Dooner H. K., Belachew A. Transposition Pattern of the Maize Element Ac from the Bz-M2(ac) Allele. Genetics. 1989 Jun;122(2):447–457. doi: 10.1093/genetics/122.2.447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Dooner H. K., Keller J., Harper E., Ralston E. Variable Patterns of Transposition of the Maize Element Activator in Tobacco. Plant Cell. 1991 May;3(5):473–482. doi: 10.1105/tpc.3.5.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Errede B., Company M., Ferchak J. D., Hutchison C. A., 3rd, Yarnell W. S. Activation regions in a yeast transposon have homology to mating type control sequences and to mammalian enhancers. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5423–5427. doi: 10.1073/pnas.82.16.5423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Flavell A. J., Smith D. B., Kumar A. Extreme heterogeneity of Ty1-copia group retrotransposons in plants. Mol Gen Genet. 1992 Jan;231(2):233–242. doi: 10.1007/BF00279796. [DOI] [PubMed] [Google Scholar]
  24. Flavell R. B. Inactivation of gene expression in plants as a consequence of specific sequence duplication. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3490–3496. doi: 10.1073/pnas.91.9.3490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Garfinkel D. J., Mastrangelo M. F., Sanders N. J., Shafer B. K., Strathern J. N. Transposon tagging using Ty elements in yeast. Genetics. 1988 Sep;120(1):95–108. doi: 10.1093/genetics/120.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Grandbastien M. A. Retroelements in higher plants. Trends Genet. 1992 Mar;8(3):103–108. doi: 10.1016/0168-9525(92)90198-d. [DOI] [PubMed] [Google Scholar]
  27. Grandbastien M. A., Spielmann A., Caboche M. Tnt1, a mobile retroviral-like transposable element of tobacco isolated by plant cell genetics. Nature. 1989 Jan 26;337(6205):376–380. doi: 10.1038/337376a0. [DOI] [PubMed] [Google Scholar]
  28. Greenblatt I. M. A chromosome replication pattern deduced from pericarp phenotypes resulting from movements of the transposable element, modulator, in maize. Genetics. 1984 Oct;108(2):471–485. doi: 10.1093/genetics/108.2.471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Haring M. A., Rommens C. M., Nijkamp H. J., Hille J. The use of transgenic plants to understand transposition mechanisms and to develop transposon tagging strategies. Plant Mol Biol. 1991 Mar;16(3):449–461. doi: 10.1007/BF00023995. [DOI] [PubMed] [Google Scholar]
  30. Hedley P. E., Machray G. C., Davies H. V., Burch L., Waugh R. cDNA cloning and expression of a potato (Solanum tuberosum) invertase. Plant Mol Biol. 1993 Aug;22(5):917–922. doi: 10.1007/BF00027378. [DOI] [PubMed] [Google Scholar]
  31. Hirochika H. Activation of tobacco retrotransposons during tissue culture. EMBO J. 1993 Jun;12(6):2521–2528. doi: 10.1002/j.1460-2075.1993.tb05907.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ichihara Y., Kurosawa Y. Construction of new T vectors for direct cloning of PCR products. Gene. 1993 Aug 16;130(1):153–154. doi: 10.1016/0378-1119(93)90361-6. [DOI] [PubMed] [Google Scholar]
  33. Ji H., Moore D. P., Blomberg M. A., Braiterman L. T., Voytas D. F., Natsoulis G., Boeke J. D. Hotspots for unselected Ty1 transposition events on yeast chromosome III are near tRNA genes and LTR sequences. Cell. 1993 Jun 4;73(5):1007–1018. doi: 10.1016/0092-8674(93)90278-x. [DOI] [PubMed] [Google Scholar]
  34. Jin Y. K., Bennetzen J. L. Structure and coding properties of Bs1, a maize retrovirus-like transposon. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6235–6239. doi: 10.1073/pnas.86.16.6235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Johns M. A., Mottinger J., Freeling M. A low copy number, copia-like transposon in maize. EMBO J. 1985 May;4(5):1093–1101. doi: 10.1002/j.1460-2075.1985.tb03745.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Jones D. A., Thomas C. M., Hammond-Kosack K. E., Balint-Kurti P. J., Jones J. D. Isolation of the tomato Cf-9 gene for resistance to Cladosporium fulvum by transposon tagging. Science. 1994 Nov 4;266(5186):789–793. doi: 10.1126/science.7973631. [DOI] [PubMed] [Google Scholar]
  37. Kaiser K., Goodwin S. F. "Site-selected" transposon mutagenesis of Drosophila. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1686–1690. doi: 10.1073/pnas.87.5.1686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Konieczny A., Voytas D. F., Cummings M. P., Ausubel F. M. A superfamily of Arabidopsis thaliana retrotransposons. Genetics. 1991 Apr;127(4):801–809. doi: 10.1093/genetics/127.4.801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Levin H. L., Weaver D. C., Boeke J. D. Novel gene expression mechanism in a fission yeast retroelement: Tf1 proteins are derived from a single primary translation product. EMBO J. 1993 Dec;12(12):4885–4895. doi: 10.1002/j.1460-2075.1993.tb06178.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Logemann J., Schell J., Willmitzer L. Improved method for the isolation of RNA from plant tissues. Anal Biochem. 1987 May 15;163(1):16–20. doi: 10.1016/0003-2697(87)90086-8. [DOI] [PubMed] [Google Scholar]
  41. Long D., Martin M., Sundberg E., Swinburne J., Puangsomlee P., Coupland G. The maize transposable element system Ac/Ds as a mutagen in Arabidopsis: identification of an albino mutation induced by Ds insertion. Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10370–10374. doi: 10.1073/pnas.90.21.10370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Manninen I., Schulman A. H. BARE-1, a copia-like retroelement in barley (Hordeum vulgare L.). Plant Mol Biol. 1993 Aug;22(5):829–846. doi: 10.1007/BF00027369. [DOI] [PubMed] [Google Scholar]
  43. Murphy G. J., Lucas H., Moore G., Flavell R. B. Sequence analysis of WIS-2-1A, a retrotransposon-like element from wheat. Plant Mol Biol. 1992 Dec;20(5):991–995. doi: 10.1007/BF00027169. [DOI] [PubMed] [Google Scholar]
  44. Pauls P. K., Kunert K., Huttner E., Grandbastien M. A. Expression of the tobacco Tnt1 retrotransposon promoter in heterologous species. Plant Mol Biol. 1994 Oct;26(1):393–402. doi: 10.1007/BF00039548. [DOI] [PubMed] [Google Scholar]
  45. Pouteau S., Huttner E., Grandbastien M. A., Caboche M. Specific expression of the tobacco Tnt1 retrotransposon in protoplasts. EMBO J. 1991 Jul;10(7):1911–1918. doi: 10.1002/j.1460-2075.1991.tb07717.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Pruitt R. E., Meyerowitz E. M. Characterization of the genome of Arabidopsis thaliana. J Mol Biol. 1986 Jan 20;187(2):169–183. doi: 10.1016/0022-2836(86)90226-3. [DOI] [PubMed] [Google Scholar]
  47. Schwebel-Dugué N., el Mtili N., Krivitzky M., Jean-Jacques I., Williams J. H., Thomas M., Kreis M., Lecharny A. Arabidopsis gene and cDNA encoding cell-wall invertase. Plant Physiol. 1994 Feb;104(2):809–810. doi: 10.1104/pp.104.2.809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Silver J., Keerikatte V. Novel use of polymerase chain reaction to amplify cellular DNA adjacent to an integrated provirus. J Virol. 1989 May;63(5):1924–1928. doi: 10.1128/jvi.63.5.1924-1928.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Smyth D. R., Kalitsis P., Joseph J. L., Sentry J. W. Plant retrotransposon from Lilium henryi is related to Ty3 of yeast and the gypsy group of Drosophila. Proc Natl Acad Sci U S A. 1989 Jul;86(13):5015–5019. doi: 10.1073/pnas.86.13.5015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Sturm A., Chrispeels M. J. cDNA cloning of carrot extracellular beta-fructosidase and its expression in response to wounding and bacterial infection. Plant Cell. 1990 Nov;2(11):1107–1119. doi: 10.1105/tpc.2.11.1107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Valvekens D., Van Montagu M., Van Lijsebettens M. Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5536–5540. doi: 10.1073/pnas.85.15.5536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Vancanneyt G., Schmidt R., O'Connor-Sanchez A., Willmitzer L., Rocha-Sosa M. Construction of an intron-containing marker gene: splicing of the intron in transgenic plants and its use in monitoring early events in Agrobacterium-mediated plant transformation. Mol Gen Genet. 1990 Jan;220(2):245–250. doi: 10.1007/BF00260489. [DOI] [PubMed] [Google Scholar]
  53. Vaucheret H., Marion-Poll A., Meyer C., Faure J. D., Marin E., Caboche M. Interest in and limits to the utilization of reporter genes for the analysis of transcriptional regulation of nitrate reductase. Mol Gen Genet. 1992 Nov;235(2-3):259–268. doi: 10.1007/BF00279369. [DOI] [PubMed] [Google Scholar]
  54. Voytas D. F., Boeke J. D. Yeast retrotransposons and tRNAs. Trends Genet. 1993 Dec;9(12):421–427. doi: 10.1016/0168-9525(93)90105-q. [DOI] [PubMed] [Google Scholar]
  55. Voytas D. F., Konieczny A., Cummings M. P., Ausubel F. M. The structure, distribution and evolution of the Ta1 retrotransposable element family of Arabidopsis thaliana. Genetics. 1990 Nov;126(3):713–721. doi: 10.1093/genetics/126.3.713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Whitham S., Dinesh-Kumar S. P., Choi D., Hehl R., Corr C., Baker B. The product of the tobacco mosaic virus resistance gene N: similarity to toll and the interleukin-1 receptor. Cell. 1994 Sep 23;78(6):1101–1115. doi: 10.1016/0092-8674(94)90283-6. [DOI] [PubMed] [Google Scholar]
  57. Xiong Y., Eickbush T. H. Origin and evolution of retroelements based upon their reverse transcriptase sequences. EMBO J. 1990 Oct;9(10):3353–3362. doi: 10.1002/j.1460-2075.1990.tb07536.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. 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]
  59. Yoshioka K., Honma H., Zushi M., Kondo S., Togashi S., Miyake T., Shiba T. Virus-like particle formation of Drosophila copia through autocatalytic processing. EMBO J. 1990 Feb;9(2):535–541. doi: 10.1002/j.1460-2075.1990.tb08140.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES