Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 1993 Mar;133(3):499–508. doi: 10.1093/genetics/133.3.499

A Ubiquitin-Conjugating Enzyme, Rad6, Affects the Distribution of Ty1 Retrotransposon Integration Positions

S W Liebman 1, G Newnam 1
PMCID: PMC1205338  PMID: 8384143

Abstract

A galactose-inducible Ty1 element was used to generate 59 independent Ty1 inserts that inactivate the CAN1 gene. As found in previous studies, the distribution of these elements shows a gradient of insertion frequency from highest to lowest between the 5' and 3' ends of the gene. However, 53 independent Ty1 and Ty2 insertions isolated by an identical procedure in an isogenic rad6 deletion strain do not show this bias. In this strain, the Ty elements insert randomly throughout CAN1. These results show that the ubiquitin-conjugating enzyme, RAD6, alters the integration site preferences of Ty1 retrotransposons.

Full Text

The Full Text of this article is available as a PDF (2.2 MB).

Selected References

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

  1. Alani E., Kleckner N. A new type of fusion analysis applicable to many organisms: protein fusions to the URA3 gene of yeast. Genetics. 1987 Sep;117(1):5–12. doi: 10.1093/genetics/117.1.5. [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., 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]
  4. 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]
  5. 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]
  6. Fink G., Farabaugh P., Roeder G., Chaleff D. Transposable elements (Ty) in yeast. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 2):575–580. doi: 10.1101/sqb.1981.045.01.074. [DOI] [PubMed] [Google Scholar]
  7. Freund R., Meselson M. Long terminal repeat nucleotide sequence and specific insertion of the gypsy transposon. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4462–4464. doi: 10.1073/pnas.81.14.4462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Haas A., Reback P. M., Pratt G., Rechsteiner M. Ubiquitin-mediated degradation of histone H3 does not require the substrate-binding ubiquitin protein ligase, E3, or attachment of polyubiquitin chains. J Biol Chem. 1990 Dec 15;265(35):21664–21669. [PubMed] [Google Scholar]
  9. Hashimoto H., Kikuchi Y., Nogi Y., Fukasawa T. Regulation of expression of the galactose gene cluster in Saccharomyces cerevisiae. Isolation and characterization of the regulatory gene GAL4. Mol Gen Genet. 1983;191(1):31–38. doi: 10.1007/BF00330886. [DOI] [PubMed] [Google Scholar]
  10. Hoffmann W. Molecular characterization of the CAN1 locus in Saccharomyces cerevisiae. A transmembrane protein without N-terminal hydrophobic signal sequence. J Biol Chem. 1985 Sep 25;260(21):11831–11837. [PubMed] [Google Scholar]
  11. 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]
  12. Jentsch S., McGrath J. P., Varshavsky A. The yeast DNA repair gene RAD6 encodes a ubiquitin-conjugating enzyme. Nature. 1987 Sep 10;329(6135):131–134. doi: 10.1038/329131a0. [DOI] [PubMed] [Google Scholar]
  13. Jentsch S., Seufert W., Hauser H. P. Genetic analysis of the ubiquitin system. Biochim Biophys Acta. 1991 Jun 13;1089(2):127–139. doi: 10.1016/0167-4781(91)90001-3. [DOI] [PubMed] [Google Scholar]
  14. Kang X. L., Yadao F., Gietz R. D., Kunz B. A. Elimination of the yeast RAD6 ubiquitin conjugase enhances base-pair transitions and G.C----T.A transversions as well as transposition of the Ty element: implications for the control of spontaneous mutation. Genetics. 1992 Feb;130(2):285–294. doi: 10.1093/genetics/130.2.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Koken M. H., Reynolds P., Jaspers-Dekker I., Prakash L., Prakash S., Bootsma D., Hoeijmakers J. H. Structural and functional conservation of two human homologs of the yeast DNA repair gene RAD6. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):8865–8869. doi: 10.1073/pnas.88.20.8865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Link A. J., Olson M. V. Physical map of the Saccharomyces cerevisiae genome at 110-kilobase resolution. Genetics. 1991 Apr;127(4):681–698. doi: 10.1093/genetics/127.4.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Louis E. J., Haber J. E. Mitotic recombination among subtelomeric Y' repeats in Saccharomyces cerevisiae. Genetics. 1990 Mar;124(3):547–559. doi: 10.1093/genetics/124.3.547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mooslehner K., Karls U., Harbers K. Retroviral integration sites in transgenic Mov mice frequently map in the vicinity of transcribed DNA regions. J Virol. 1990 Jun;64(6):3056–3058. doi: 10.1128/jvi.64.6.3056-3058.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Morrison A., Miller E. J., Prakash L. Domain structure and functional analysis of the carboxyl-terminal polyacidic sequence of the RAD6 protein of Saccharomyces cerevisiae. Mol Cell Biol. 1988 Mar;8(3):1179–1185. doi: 10.1128/mcb.8.3.1179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Oyen T. B., Gabrielsen O. S. Non-random distribution of the Ty1 elements within nuclear DNA of Saccharomyces cerevisiae. FEBS Lett. 1983 Sep 19;161(2):201–206. doi: 10.1016/0014-5793(83)81007-2. [DOI] [PubMed] [Google Scholar]
  22. Picologlou S., Brown N., Liebman S. W. Mutations in RAD6, a yeast gene encoding a ubiquitin-conjugating enzyme, stimulate retrotransposition. Mol Cell Biol. 1990 Mar;10(3):1017–1022. doi: 10.1128/mcb.10.3.1017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pryciak P. M., Varmus H. E. Nucleosomes, DNA-binding proteins, and DNA sequence modulate retroviral integration target site selection. Cell. 1992 May 29;69(5):769–780. doi: 10.1016/0092-8674(92)90289-o. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  27. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  28. Sandmeyer S. B., Hansen L. J., Chalker D. L. Integration specificity of retrotransposons and retroviruses. Annu Rev Genet. 1990;24:491–518. doi: 10.1146/annurev.ge.24.120190.002423. [DOI] [PubMed] [Google Scholar]
  29. Sathe G. M., O'Brien S., McLaughlin M. M., Watson F., Livi G. P. Use of polymerase chain reaction for rapid detection of gene insertions in whole yeast cells. Nucleic Acids Res. 1991 Sep 11;19(17):4775–4775. doi: 10.1093/nar/19.17.4775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Scherdin U., Rhodes K., Breindl M. Transcriptionally active genome regions are preferred targets for retrovirus integration. J Virol. 1990 Feb;64(2):907–912. doi: 10.1128/jvi.64.2.907-912.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Shimotohno K., Temin H. M. No apparent nucleotide sequence specificity in cellular DNA juxtaposed to retrovirus proviruses. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7357–7361. doi: 10.1073/pnas.77.12.7357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Simchen G., Winston F., Styles C. A., Fink G. R. Ty-mediated gene expression of the LYS2 and HIS4 genes of Saccharomyces cerevisiae is controlled by the same SPT genes. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2431–2434. doi: 10.1073/pnas.81.8.2431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Snyder M. P., Kimbrell D., Hunkapiller M., Hill R., Fristrom J., Davidson N. A transposable element that splits the promoter region inactivates a Drosophila cuticle protein gene. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7430–7434. doi: 10.1073/pnas.79.23.7430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sung P., Prakash S., Prakash L. The RAD6 protein of Saccharomyces cerevisiae polyubiquitinates histones, and its acidic domain mediates this activity. Genes Dev. 1988 Nov;2(11):1476–1485. doi: 10.1101/gad.2.11.1476. [DOI] [PubMed] [Google Scholar]
  36. Swerdlow P. S., Schuster T., Finley D. A conserved sequence in histone H2A which is a ubiquitination site in higher eucaryotes is not required for growth in Saccharomyces cerevisiae. Mol Cell Biol. 1990 Sep;10(9):4905–4911. doi: 10.1128/mcb.10.9.4905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. 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]
  38. 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]
  39. Warmington J. R., Anwar R., Newlon C. S., Waring R. B., Davies R. W., Indge K. J., Oliver S. G. A 'hot-spot' for Ty transposition on the left arm of yeast chromosome III. Nucleic Acids Res. 1986 Apr 25;14(8):3475–3485. doi: 10.1093/nar/14.8.3475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Weinstock K. G., Mastrangelo M. F., Burkett T. J., Garfinkel D. J., Strathern J. N. Multimeric arrays of the yeast retrotransposon Ty. Mol Cell Biol. 1990 Jun;10(6):2882–2892. doi: 10.1128/mcb.10.6.2882. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Whelan W. L., Gocke E., Manney T. R. The CAN1 locus of Saccharomyces cerevisiae: fine-structure analysis and forward mutation rates. Genetics. 1979 Jan;91(1):35–51. doi: 10.1093/genetics/91.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. 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]
  43. Williamson V. M., Cox D., Young E. T., Russell D. W., Smith M. Characterization of transposable element-associated mutations that alter yeast alcohol dehydrogenase II expression. Mol Cell Biol. 1983 Jan;3(1):20–31. doi: 10.1128/mcb.3.1.20. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Winston F., Chumley F., Fink G. R. Eviction and transplacement of mutant genes in yeast. Methods Enzymol. 1983;101:211–228. doi: 10.1016/0076-6879(83)01016-2. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES