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. 2003 Sep;165(1):47–63. doi: 10.1093/genetics/165.1.47

Patterns of heteroduplex formation associated with the initiation of meiotic recombination in the yeast Saccharomyces cerevisiae.

Jason D Merker 1, Margaret Dominska 1, Thomas D Petes 1
PMCID: PMC1462766  PMID: 14504217

Abstract

The double-strand break repair (DSBR) model of recombination predicts that heteroduplexes will be formed in regions that flank the double-strand break (DSB) site and that the resulting intermediate is resolved to generate either crossovers or noncrossovers for flanking markers. Previous studies in Saccharomyces cerevisiae, however, failed to detect heteroduplexes on both sides of the DSB site. Recent physical studies suggest that some recombination events involve heterodupex formation by a mechanism, synthesis-dependent strand annealing (SDSA), that is inherently asymmetric with respect to the DSB site and that leads exclusively to noncrossovers of flanking markers. Below, we demonstrate that many of the recombination events initiated at the HIS4 recombination hotspot are consistent with a variant of the DSBR model in which the extent of heteroduplex on one side of the DSB site is much greater than that on the other. Events that include only one flanking marker in the heteroduplex (unidirectional events) are usually resolved as noncrossovers, whereas events that include both flanking markers (bidirectional events) are usually resolved as crossovers. The unidirectional events may represent SDSA, consistent with the conclusions of others, although other possibilities are not excluded. We also show that the level of recombination reflects the integration of events initiated at several different DSB sites, and we identify a subset of gene conversion events that may involve break-induced replication (BIR) or repair of a double-stranded DNA gap.

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Selected References

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  1. Alani E., Padmore R., Kleckner N. Analysis of wild-type and rad50 mutants of yeast suggests an intimate relationship between meiotic chromosome synapsis and recombination. Cell. 1990 May 4;61(3):419–436. doi: 10.1016/0092-8674(90)90524-i. [DOI] [PubMed] [Google Scholar]
  2. Alani E., Reenan R. A., Kolodner R. D. Interaction between mismatch repair and genetic recombination in Saccharomyces cerevisiae. Genetics. 1994 May;137(1):19–39. doi: 10.1093/genetics/137.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Alani E. The Saccharomyces cerevisiae Msh2 and Msh6 proteins form a complex that specifically binds to duplex oligonucleotides containing mismatched DNA base pairs. Mol Cell Biol. 1996 Oct;16(10):5604–5615. doi: 10.1128/mcb.16.10.5604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Allers T., Lichten M. Differential timing and control of noncrossover and crossover recombination during meiosis. Cell. 2001 Jul 13;106(1):47–57. doi: 10.1016/s0092-8674(01)00416-0. [DOI] [PubMed] [Google Scholar]
  5. Allers T., Lichten M. Intermediates of yeast meiotic recombination contain heteroduplex DNA. Mol Cell. 2001 Jul;8(1):225–231. doi: 10.1016/s1097-2765(01)00280-5. [DOI] [PubMed] [Google Scholar]
  6. Baudat F., Nicolas A. Clustering of meiotic double-strand breaks on yeast chromosome III. Proc Natl Acad Sci U S A. 1997 May 13;94(10):5213–5218. doi: 10.1073/pnas.94.10.5213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bergerat A., de Massy B., Gadelle D., Varoutas P. C., Nicolas A., Forterre P. An atypical topoisomerase II from Archaea with implications for meiotic recombination. Nature. 1997 Mar 27;386(6623):414–417. doi: 10.1038/386414a0. [DOI] [PubMed] [Google Scholar]
  8. Borde V., Wu T. C., Lichten M. Use of a recombination reporter insert to define meiotic recombination domains on chromosome III of Saccharomyces cerevisiae. Mol Cell Biol. 1999 Jul;19(7):4832–4842. doi: 10.1128/mcb.19.7.4832. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Detloff P., Petes T. D. Measurements of excision repair tracts formed during meiotic recombination in Saccharomyces cerevisiae. Mol Cell Biol. 1992 Apr;12(4):1805–1814. doi: 10.1128/mcb.12.4.1805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Detloff P., White M. A., Petes T. D. Analysis of a gene conversion gradient at the HIS4 locus in Saccharomyces cerevisiae. Genetics. 1992 Sep;132(1):113–123. doi: 10.1093/genetics/132.1.113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fan Q., Xu F., Petes T. D. Meiosis-specific double-strand DNA breaks at the HIS4 recombination hot spot in the yeast Saccharomyces cerevisiae: control in cis and trans. Mol Cell Biol. 1995 Mar;15(3):1679–1688. doi: 10.1128/mcb.15.3.1679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Foss H. M., Hillers K. J., Stahl F. W. The conversion gradient at HIS4 of Saccharomyces cerevisiae. II. A role for mismatch repair directed by biased resolution of the recombinational intermediate. Genetics. 1999 Oct;153(2):573–583. doi: 10.1093/genetics/153.2.573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gerton J. L., DeRisi J., Shroff R., Lichten M., Brown P. O., Petes T. D. Global mapping of meiotic recombination hotspots and coldspots in the yeast Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 2000 Oct 10;97(21):11383–11390. doi: 10.1073/pnas.97.21.11383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gilbertson L. A., Stahl F. W. A test of the double-strand break repair model for meiotic recombination in Saccharomyces cerevisiae. Genetics. 1996 Sep;144(1):27–41. doi: 10.1093/genetics/144.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hillers K. J., Stahl F. W. The conversion gradient at HIS4 of Saccharomyces cerevisiae. I. Heteroduplex rejection and restoration of Mendelian segregation. Genetics. 1999 Oct;153(2):555–572. doi: 10.1093/genetics/153.2.555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Keeney S., Giroux C. N., Kleckner N. Meiosis-specific DNA double-strand breaks are catalyzed by Spo11, a member of a widely conserved protein family. Cell. 1997 Feb 7;88(3):375–384. doi: 10.1016/s0092-8674(00)81876-0. [DOI] [PubMed] [Google Scholar]
  17. Kirkpatrick D. T., Fan Q., Petes T. D. Maximal stimulation of meiotic recombination by a yeast transcription factor requires the transcription activation domain and a DNA-binding domain. Genetics. 1999 May;152(1):101–115. doi: 10.1093/genetics/152.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nag D. K., Kurst A. A 140-bp-long palindromic sequence induces double-strand breaks during meiosis in the yeast Saccharomyces cerevisiae. Genetics. 1997 Jul;146(3):835–847. doi: 10.1093/genetics/146.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nag D. K., Petes T. D. Genetic evidence for preferential strand transfer during meiotic recombination in yeast. Genetics. 1990 Aug;125(4):753–761. doi: 10.1093/genetics/125.4.753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nag D. K., Petes T. D. Physical detection of heteroduplexes during meiotic recombination in the yeast Saccharomyces cerevisiae. Mol Cell Biol. 1993 Apr;13(4):2324–2331. doi: 10.1128/mcb.13.4.2324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nag D. K., Petes T. D. Seven-base-pair inverted repeats in DNA form stable hairpins in vivo in Saccharomyces cerevisiae. Genetics. 1991 Nov;129(3):669–673. doi: 10.1093/genetics/129.3.669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nag D. K., White M. A., Petes T. D. Palindromic sequences in heteroduplex DNA inhibit mismatch repair in yeast. Nature. 1989 Jul 27;340(6231):318–320. doi: 10.1038/340318a0. [DOI] [PubMed] [Google Scholar]
  23. Perkins D. D. Biochemical Mutants in the Smut Fungus Ustilago Maydis. Genetics. 1949 Sep;34(5):607–626. doi: 10.1093/genetics/34.5.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Petes Thomas D., Merker Jason D. Context dependence of meiotic recombination hotspots in yeast: the relationship between recombination activity of a reporter construct and base composition. Genetics. 2002 Dec;162(4):2049–2052. doi: 10.1093/genetics/162.4.2049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Porter S. E., White M. A., Petes T. D. Genetic evidence that the meiotic recombination hotspot at the HIS4 locus of Saccharomyces cerevisiae does not represent a site for a symmetrically processed double-strand break. Genetics. 1993 May;134(1):5–19. doi: 10.1093/genetics/134.1.5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pâques F., Haber J. E. Multiple pathways of recombination induced by double-strand breaks in Saccharomyces cerevisiae. Microbiol Mol Biol Rev. 1999 Jun;63(2):349–404. doi: 10.1128/mmbr.63.2.349-404.1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schwacha A., Kleckner N. Identification of double Holliday junctions as intermediates in meiotic recombination. Cell. 1995 Dec 1;83(5):783–791. doi: 10.1016/0092-8674(95)90191-4. [DOI] [PubMed] [Google Scholar]
  28. Stahl F. W., Hillers K. J. Heteroduplex rejection in yeast? Genetics. 2000 Apr;154(4):1913–1916. doi: 10.1093/genetics/154.4.1913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stapleton A., Petes T. D. The Tn3 beta-lactamase gene acts as a hotspot for meiotic recombination in yeast. Genetics. 1991 Jan;127(1):39–51. doi: 10.1093/genetics/127.1.39. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Sun H., Treco D., Schultes N. P., Szostak J. W. Double-strand breaks at an initiation site for meiotic gene conversion. Nature. 1989 Mar 2;338(6210):87–90. doi: 10.1038/338087a0. [DOI] [PubMed] [Google Scholar]
  32. Sun H., Treco D., Szostak J. W. Extensive 3'-overhanging, single-stranded DNA associated with the meiosis-specific double-strand breaks at the ARG4 recombination initiation site. Cell. 1991 Mar 22;64(6):1155–1161. doi: 10.1016/0092-8674(91)90270-9. [DOI] [PubMed] [Google Scholar]
  33. Symington L. S., Petes T. D. Expansions and contractions of the genetic map relative to the physical map of yeast chromosome III. Mol Cell Biol. 1988 Feb;8(2):595–604. doi: 10.1128/mcb.8.2.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Szostak J. W., Orr-Weaver T. L., Rothstein R. J., Stahl F. W. The double-strand-break repair model for recombination. Cell. 1983 May;33(1):25–35. doi: 10.1016/0092-8674(83)90331-8. [DOI] [PubMed] [Google Scholar]
  35. Wach A., Brachat A., Pöhlmann R., Philippsen P. New heterologous modules for classical or PCR-based gene disruptions in Saccharomyces cerevisiae. Yeast. 1994 Dec;10(13):1793–1808. doi: 10.1002/yea.320101310. [DOI] [PubMed] [Google Scholar]
  36. White M. A., Petes T. D. Analysis of meiotic recombination events near a recombination hotspot in the yeast Saccharomyces cerevisiae. Curr Genet. 1994 Jul;26(1):21–30. doi: 10.1007/BF00326300. [DOI] [PubMed] [Google Scholar]

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