Skip to main content
Genetics logoLink to Genetics
. 1994 Dec;138(4):1055–1065. doi: 10.1093/genetics/138.4.1055

An Implanted Recombination Hot Spot Stimulates Recombination and Enhances Sister Chromatid Cohesion of Heterologous Yacs during Yeast Meiosis

D D Sears 1, P Hieter 1, G Simchen 1
PMCID: PMC1206247  PMID: 7896090

Abstract

Heterologous yeast artificial chromosomes (YACs) do not recombine with each other and missegregate in 25% of meiosis I events. Recombination hot spots in the yeast Saccharomyces cerevisiae have previously been shown to be associated with sites of meiosis-induced double-strand breaks (DSBs). A 6-kb fragment containing a recombination hot spot/DSB site was implanted onto two heterologous human DNA YACs and was shown to cause the YACs to undergo meiotic recombination in 5-8% of tetrads. Reciprocal exchanges initiated and resolved within the 6-kb insert. Presence of the insert had no detectable effect on meiosis I nondisjunction. Surprisingly, the recombination hot spots acted in cis to significantly reduce precocious sister-chromatid segregation. This novel observation suggests that DSBs are instrumental in maintaining cohesion between sister chromatids in meiosis I. We propose that this previously unknown function of DSBs is mediated by the stimulation of sister-chromatid exchange and/or its intermediates.

Full Text

The Full Text of this article is available as a PDF (5.7 MB).

Selected References

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

  1. Bentley D. R., Todd C., Collins J., Holland J., Dunham I., Hassock S., Bankier A., Giannelli F. The development and application of automated gridding for efficient screening of yeast and bacterial ordered libraries. Genomics. 1992 Mar;12(3):534–541. doi: 10.1016/0888-7543(92)90445-x. [DOI] [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. Cao L., Alani E., Kleckner N. A pathway for generation and processing of double-strand breaks during meiotic recombination in S. cerevisiae. Cell. 1990 Jun 15;61(6):1089–1101. doi: 10.1016/0092-8674(90)90072-m. [DOI] [PubMed] [Google Scholar]
  4. Carle G. F., Olson M. V. Separation of chromosomal DNA molecules from yeast by orthogonal-field-alternation gel electrophoresis. Nucleic Acids Res. 1984 Jul 25;12(14):5647–5664. doi: 10.1093/nar/12.14.5647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carpenter A. T. Distributive segregation: motors in the polar wind? Cell. 1991 Mar 8;64(5):885–890. doi: 10.1016/0092-8674(91)90313-n. [DOI] [PubMed] [Google Scholar]
  6. Chattoo B. B., Sherman F., Azubalis D. A., Fjellstedt T. A., Mehnert D., Ogur M. Selection of lys2 Mutants of the Yeast SACCHAROMYCES CEREVISIAE by the Utilization of alpha-AMINOADIPATE. Genetics. 1979 Sep;93(1):51–65. doi: 10.1093/genetics/93.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Collins I., Newlon C. S. Meiosis-specific formation of joint DNA molecules containing sequences from homologous chromosomes. Cell. 1994 Jan 14;76(1):65–75. doi: 10.1016/0092-8674(94)90173-2. [DOI] [PubMed] [Google Scholar]
  8. Dawson D. S., Murray A. W., Szostak J. W. An alternative pathway for meiotic chromosome segregation in yeast. Science. 1986 Nov 7;234(4777):713–717. doi: 10.1126/science.3535068. [DOI] [PubMed] [Google Scholar]
  9. Game J. C. Pulsed-field gel analysis of the pattern of DNA double-strand breaks in the Saccharomyces genome during meiosis. Dev Genet. 1992;13(6):485–497. doi: 10.1002/dvg.1020130610. [DOI] [PubMed] [Google Scholar]
  10. Game J. C., Sitney K. C., Cook V. E., Mortimer R. K. Use of a ring chromosome and pulsed-field gels to study interhomolog recombination, double-strand DNA breaks and sister-chromatid exchange in yeast. Genetics. 1989 Dec;123(4):695–713. doi: 10.1093/genetics/123.4.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goldway M., Sherman A., Zenvirth D., Arbel T., Simchen G. A short chromosomal region with major roles in yeast chromosome III meiotic disjunction, recombination and double strand breaks. Genetics. 1993 Feb;133(2):159–169. doi: 10.1093/genetics/133.2.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Guacci V., Kaback D. B. Distributive disjunction of authentic chromosomes in Saccharomyces cerevisiae. Genetics. 1991 Mar;127(3):475–488. doi: 10.1093/genetics/127.3.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Haber J. E., Thorburn P. C., Rogers D. Meiotic and mitotic behavior of dicentric chromosomes in Saccharomyces cerevisiae. Genetics. 1984 Feb;106(2):185–205. doi: 10.1093/genetics/106.2.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hawley R. S., Arbel T. Yeast genetics and the fall of the classical view of meiosis. Cell. 1993 Feb 12;72(3):301–303. doi: 10.1016/0092-8674(93)90108-3. [DOI] [PubMed] [Google Scholar]
  15. Hawley R. S., McKim K. S., Arbel T. Meiotic segregation in Drosophila melanogaster females: molecules, mechanisms, and myths. Annu Rev Genet. 1993;27:281–317. doi: 10.1146/annurev.ge.27.120193.001433. [DOI] [PubMed] [Google Scholar]
  16. Jackson J. A., Fink G. R. Meiotic recombination between duplicated genetic elements in Saccharomyces cerevisiae. Genetics. 1985 Feb;109(2):303–332. doi: 10.1093/genetics/109.2.303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kaback D. B. Meiotic segregation of circular plasmid-minichromosomes from intact chromosomes in Saccharomyces cerevisiae. Curr Genet. 1989 Jun;15(6):385–392. doi: 10.1007/BF00376792. [DOI] [PubMed] [Google Scholar]
  18. Kleckner N., Padmore R., Bishop D. K. Meiotic chromosome metabolism: one view. Cold Spring Harb Symp Quant Biol. 1991;56:729–743. doi: 10.1101/sqb.1991.056.01.082. [DOI] [PubMed] [Google Scholar]
  19. Lambie E. J., Roeder G. S. A yeast centromere acts in cis to inhibit meiotic gene conversion of adjacent sequences. Cell. 1988 Mar 25;52(6):863–873. doi: 10.1016/0092-8674(88)90428-x. [DOI] [PubMed] [Google Scholar]
  20. Lambie E. J., Roeder G. S. Repression of meiotic crossing over by a centromere (CEN3) in Saccharomyces cerevisiae. Genetics. 1986 Nov;114(3):769–789. doi: 10.1093/genetics/114.3.769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Loidl J., Scherthan H., Kaback D. B. Physical association between nonhomologous chromosomes precedes distributive disjunction in yeast. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):331–334. doi: 10.1073/pnas.91.1.331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Maguire M. P. Sister chromatid cohesiveness: vital function, obscure mechanism. Biochem Cell Biol. 1990 Nov;68(11):1231–1242. doi: 10.1139/o90-183. [DOI] [PubMed] [Google Scholar]
  23. Mann C., Davis R. W. Meiotic disjunction of circular minichromosomes in yeast does not require DNA homology. Proc Natl Acad Sci U S A. 1986 Aug;83(16):6017–6019. doi: 10.1073/pnas.83.16.6017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McKim K. S., Peters K., Rose A. M. Two types of sites required for meiotic chromosome pairing in Caenorhabditis elegans. Genetics. 1993 Jul;134(3):749–768. doi: 10.1093/genetics/134.3.749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mortimer R. K., Schild D., Contopoulou C. R., Kans J. A. Genetic map of Saccharomyces cerevisiae, edition 10. Yeast. 1989 Sep-Oct;5(5):321–403. doi: 10.1002/yea.320050503. [DOI] [PubMed] [Google Scholar]
  26. Murray A. W., Szostak J. W. Chromosome segregation in mitosis and meiosis. Annu Rev Cell Biol. 1985;1:289–315. doi: 10.1146/annurev.cb.01.110185.001445. [DOI] [PubMed] [Google Scholar]
  27. Pavan W. J., Hieter P., Reeves R. H. Generation of deletion derivatives by targeted transformation of human-derived yeast artificial chromosomes. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1300–1304. doi: 10.1073/pnas.87.4.1300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Roeder G. S. Chromosome synapsis and genetic recombination: their roles in meiotic chromosome segregation. Trends Genet. 1990 Dec;6(12):385–389. doi: 10.1016/0168-9525(90)90297-j. [DOI] [PubMed] [Google Scholar]
  29. Ross L. O., Treco D., Nicolas A., Szostak J. W., Dawson D. Meiotic recombination on artificial chromosomes in yeast. Genetics. 1992 Jul;131(3):541–550. doi: 10.1093/genetics/131.3.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schwacha A., Kleckner N. Identification of joint molecules that form frequently between homologs but rarely between sister chromatids during yeast meiosis. Cell. 1994 Jan 14;76(1):51–63. doi: 10.1016/0092-8674(94)90172-4. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Suja J. A., Antonio C., Rufas J. S. Involvement of chromatid cohesiveness at the centromere and chromosome arms in meiotic chromosome segregation: a cytological approach. Chromosoma. 1992 Jun;101(8):493–501. doi: 10.1007/BF00352472. [DOI] [PubMed] [Google Scholar]
  33. Sun H., Dawson D., Szostak J. W. Genetic and physical analyses of sister chromatid exchange in yeast meiosis. Mol Cell Biol. 1991 Dec;11(12):6328–6336. doi: 10.1128/mcb.11.12.6328. [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. Vollrath D., Davis R. W., Connelly C., Hieter P. Physical mapping of large DNA by chromosome fragmentation. Proc Natl Acad Sci U S A. 1988 Aug;85(16):6027–6031. doi: 10.1073/pnas.85.16.6027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wagstaff J. E., Klapholz S., Waddell C. S., Jensen L., Esposito R. E. Meiotic exchange within and between chromosomes requires a common Rec function in Saccharomyces cerevisiae. Mol Cell Biol. 1985 Dec;5(12):3532–3544. doi: 10.1128/mcb.5.12.3532. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wu T. C., Lichten M. Meiosis-induced double-strand break sites determined by yeast chromatin structure. Science. 1994 Jan 28;263(5146):515–518. doi: 10.1126/science.8290959. [DOI] [PubMed] [Google Scholar]
  38. Zenvirth D., Arbel T., Sherman A., Goldway M., Klein S., Simchen G. Multiple sites for double-strand breaks in whole meiotic chromosomes of Saccharomyces cerevisiae. EMBO J. 1992 Sep;11(9):3441–3447. doi: 10.1002/j.1460-2075.1992.tb05423.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES