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. 1994 May;137(1):41–48. doi: 10.1093/genetics/137.1.41

Plasmid-Mediated Induction of Recombination in Yeast

R Silberman 1, M Kupiec 1
PMCID: PMC1205951  PMID: 8056321

Abstract

Diploid yeast cells heteroallelic at the HIS3 locus were transformed with a minichromosome (centromeric plasmid) carrying homology to the HIS3 region and containing the same two mutations as were present in the chromosomes. When a double-strand break (DSB) was introduced in the region of homology, an increase in the recombination frequency between heteroalleles (leading to His(+) cells) was observed, although the plasmid was unable to donate wild-type information. This induction of recombination was dependent on the presence of homology between the plasmid sequences and the chromosomes. We show evidence for the physical involvement of the plasmid in tripartite recombination events, and we propose models that can explain the interactions between the plasmid-borne and chromosomal-borne alleles. Our results suggest that the mitotic induction of recombination by DNA damage is due to localized initiation of recombination events, and not to a general induction of recombination enzymes in the cell.

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Selected References

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

  1. Ahn B. Y., Dornfeld K. J., Fagrelius T. J., Livingston D. M. Effect of limited homology on gene conversion in a Saccharomyces cerevisiae plasmid recombination system. Mol Cell Biol. 1988 Jun;8(6):2442–2448. doi: 10.1128/mcb.8.6.2442-2448.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Fabre F., Roman H. Genetic evidence for inducibility of recombination competence in yeast. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1667–1671. doi: 10.1073/pnas.74.4.1667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gietz R. D., Sugino A. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene. 1988 Dec 30;74(2):527–534. doi: 10.1016/0378-1119(88)90185-0. [DOI] [PubMed] [Google Scholar]
  5. Hanawalt P. C., Cooper P. K., Ganesan A. K., Smith C. A. DNA repair in bacteria and mammalian cells. Annu Rev Biochem. 1979;48:783–836. doi: 10.1146/annurev.bi.48.070179.004031. [DOI] [PubMed] [Google Scholar]
  6. Lee M. G., Yarranton G. T. Inducible DNA repair in Ustilago maydis. Mol Gen Genet. 1982;185(2):245–250. doi: 10.1007/BF00330793. [DOI] [PubMed] [Google Scholar]
  7. Little J. W., Mount D. W. The SOS regulatory system of Escherichia coli. Cell. 1982 May;29(1):11–22. doi: 10.1016/0092-8674(82)90085-x. [DOI] [PubMed] [Google Scholar]
  8. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. Yeast transformation: a model system for the study of recombination. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6354–6358. doi: 10.1073/pnas.78.10.6354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Perera J. R., Glasunov A. V., Glaser V. M., Boreiko A. V. Repair of double-strand breaks in plasmid DNA in the yeast Saccharomyces cerevisiae. Mol Gen Genet. 1988 Aug;213(2-3):421–424. doi: 10.1007/BF00339611. [DOI] [PubMed] [Google Scholar]
  10. ROMAN H., JACOB F. A comparison of spontaneous and ultraviolet-induced allelic recombination with reference to the recombination of outside markers. Cold Spring Harb Symp Quant Biol. 1958;23:155–160. doi: 10.1101/sqb.1958.023.01.019. [DOI] [PubMed] [Google Scholar]
  11. Ray A., Machin N., Stahl F. W. A DNA double chain break stimulates triparental recombination in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1989 Aug;86(16):6225–6229. doi: 10.1073/pnas.86.16.6225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Roitgrund C., Steinlauf R., Kupiec M. Donation of information to the unbroken chromosome in double-strand break repair. Curr Genet. 1993 May-Jun;23(5-6):414–422. doi: 10.1007/BF00312628. [DOI] [PubMed] [Google Scholar]
  13. Schiestl R. H., Gietz R. D. High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier. Curr Genet. 1989 Dec;16(5-6):339–346. doi: 10.1007/BF00340712. [DOI] [PubMed] [Google Scholar]
  14. Schiestl R. H., Petes T. D. Integration of DNA fragments by illegitimate recombination in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7585–7589. doi: 10.1073/pnas.88.17.7585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Simon J. R., Moore P. D. Induction of homologous recombination in Saccharomyces cerevisiae. Mol Gen Genet. 1988 Sep;214(1):37–41. doi: 10.1007/BF00340176. [DOI] [PubMed] [Google Scholar]
  16. WILKIE D., LEWIS D. THE EFFECT OF ULTRAVIOLET LIGHT ON RECOMBINATION IN YEAST. Genetics. 1963 Dec;48:1701–1716. doi: 10.1093/genetics/48.12.1701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Zgaga Z., Chanet R., Radman M., Fabre F. Mismatch-stimulated plasmid integration in yeast. Curr Genet. 1991 Apr;19(4):329–332. doi: 10.1007/BF00355064. [DOI] [PubMed] [Google Scholar]

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