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. 1992 Sep;11(9):3441–3447. doi: 10.1002/j.1460-2075.1992.tb05423.x

Multiple sites for double-strand breaks in whole meiotic chromosomes of Saccharomyces cerevisiae.

D Zenvirth 1, T Arbel 1, A Sherman 1, M Goldway 1, S Klein 1, G Simchen 1
PMCID: PMC556879  PMID: 1324174

Abstract

We present a scheme for locating double-strand breaks (DSBs) in meiotic chromosomes of Saccharomyces cerevisiae, based on the separation of large DNA molecules by pulsed field gel electrophoresis. Using a rad50S mutant, in which DSBs are not processed, we show that DSBs are widely induced in S. cerevisiae chromosomes during meiosis. Some of the DSBs accumulate at certain preferred sites. We present general profiles of DSBs in chromosomes III, V, VI and VII. A map of the 12 preferred sites on chromosome III is presented. At least some of these sites correlate with known 'hot spots' for meiotic recombination. The data are discussed in view of current models of meiotic recombination and chromosome segregation.

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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. Broach J. R., Strathern J. N., Hicks J. B. Transformation in yeast: development of a hybrid cloning vector and isolation of the CAN1 gene. Gene. 1979 Dec;8(1):121–133. doi: 10.1016/0378-1119(79)90012-x. [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. Carpenter A. T. Gene conversion, recombination nodules, and the initiation of meiotic synapsis. Bioessays. 1987 May;6(5):232–236. doi: 10.1002/bies.950060510. [DOI] [PubMed] [Google Scholar]
  5. Chanet R., Magana-Schwencke N., Fabre F. Potential DNA-binding domains in the RAD18 gene product of Saccharomyces cerevisiae. Gene. 1988 Dec 30;74(2):543–547. doi: 10.1016/0378-1119(88)90187-4. [DOI] [PubMed] [Google Scholar]
  6. Clarke L., Carbon J. Isolation of a yeast centromere and construction of functional small circular chromosomes. Nature. 1980 Oct 9;287(5782):504–509. doi: 10.1038/287504a0. [DOI] [PubMed] [Google Scholar]
  7. Detloff P., Sieber J., Petes T. D. Repair of specific base pair mismatches formed during meiotic recombination in the yeast Saccharomyces cerevisiae. Mol Cell Biol. 1991 Feb;11(2):737–745. doi: 10.1128/mcb.11.2.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Donahue T. F., Farabaugh P. J., Fink G. R. The nucleotide sequence of the HIS4 region of yeast. Gene. 1982 Apr;18(1):47–59. doi: 10.1016/0378-1119(82)90055-5. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Gerring S. L., Connelly C., Hieter P. Positional mapping of genes by chromosome blotting and chromosome fragmentation. Methods Enzymol. 1991;194:57–77. doi: 10.1016/0076-6879(91)94007-y. [DOI] [PubMed] [Google Scholar]
  11. Higgins D. R., Prakash L., Reynolds P., Prakash S. Isolation and characterization of the RAD2 gene of Saccharomyces cerevisiae. Gene. 1984 Oct;30(1-3):121–128. doi: 10.1016/0378-1119(84)90112-4. [DOI] [PubMed] [Google Scholar]
  12. Hitzeman R. A., Clarke L., Carbon J. Isolation and characterization of the yeast 3-phosphoglycerokinase gene (PGK) by an immunological screening technique. J Biol Chem. 1980 Dec 25;255(24):12073–12080. [PubMed] [Google Scholar]
  13. Hsu Y. P., Schimmel P. Yeast LEU1. Repression of mRNA levels by leucine and relationship of 5'-noncoding region to that of LEU2. J Biol Chem. 1984 Mar 25;259(6):3714–3719. [PubMed] [Google Scholar]
  14. Kassir Y., Simchen G. Monitoring meiosis and sporulation in Saccharomyces cerevisiae. Methods Enzymol. 1991;194:94–110. doi: 10.1016/0076-6879(91)94009-2. [DOI] [PubMed] [Google Scholar]
  15. Klar A. J., Miglio L. M. Initiation of meiotic recombination by double-strand DNA breaks in S. pombe. Cell. 1986 Aug 29;46(5):725–731. doi: 10.1016/0092-8674(86)90348-x. [DOI] [PubMed] [Google Scholar]
  16. Kolodkin A. L., Klar A. J., Stahl F. W. Double-strand breaks can initiate meiotic recombination in S. cerevisiae. Cell. 1986 Aug 29;46(5):733–740. doi: 10.1016/0092-8674(86)90349-1. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Ma H., Kunes S., Schatz P. J., Botstein D. Plasmid construction by homologous recombination in yeast. Gene. 1987;58(2-3):201–216. doi: 10.1016/0378-1119(87)90376-3. [DOI] [PubMed] [Google Scholar]
  19. Maguire M. P. Homologous chromosome pairing. Philos Trans R Soc Lond B Biol Sci. 1977 Mar 21;277(955):245–258. doi: 10.1098/rstb.1977.0015. [DOI] [PubMed] [Google Scholar]
  20. Meselson M. S., Radding C. M. A general model for genetic recombination. Proc Natl Acad Sci U S A. 1975 Jan;72(1):358–361. doi: 10.1073/pnas.72.1.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Montelone B. A., Hoekstra M. F., Malone R. E. Spontaneous mitotic recombination in yeast: the hyper-recombinational rem1 mutations are alleles of the RAD3 gene. Genetics. 1988 Jun;119(2):289–301. doi: 10.1093/genetics/119.2.289. [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. Neff N. F., Thomas J. H., Grisafi P., Botstein D. Isolation of the beta-tubulin gene from yeast and demonstration of its essential function in vivo. Cell. 1983 May;33(1):211–219. doi: 10.1016/0092-8674(83)90350-1. [DOI] [PubMed] [Google Scholar]
  24. Newlon C. S., Lipchitz L. R., Collins I., Deshpande A., Devenish R. J., Green R. P., Klein H. L., Palzkill T. G., Ren R. B., Synn S. Analysis of a circular derivative of Saccharomyces cerevisiae chromosome III: a physical map and identification and location of ARS elements. Genetics. 1991 Oct;129(2):343–357. doi: 10.1093/genetics/129.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Oliver S. G., van der Aart Q. J., Agostoni-Carbone M. L., Aigle M., Alberghina L., Alexandraki D., Antoine G., Anwar R., Ballesta J. P., Benit P. The complete DNA sequence of yeast chromosome III. Nature. 1992 May 7;357(6373):38–46. doi: 10.1038/357038a0. [DOI] [PubMed] [Google Scholar]
  26. Orr-Weaver T. L., Szostak J. W. Fungal recombination. Microbiol Rev. 1985 Mar;49(1):33–58. doi: 10.1128/mr.49.1.33-58.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Padmore R., Cao L., Kleckner N. Temporal comparison of recombination and synaptonemal complex formation during meiosis in S. cerevisiae. Cell. 1991 Sep 20;66(6):1239–1256. doi: 10.1016/0092-8674(91)90046-2. [DOI] [PubMed] [Google Scholar]
  29. Paquin C. E., Williamson V. M. Ty insertions at two loci account for most of the spontaneous antimycin A resistance mutations during growth at 15 degrees C of Saccharomyces cerevisiae strains lacking ADH1. Mol Cell Biol. 1986 Jan;6(1):70–79. doi: 10.1128/mcb.6.1.70. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ray B. L., White C. I., Haber J. E. The TSM1 gene of Saccharomyces cerevisiae overlaps the MAT locus. Curr Genet. 1991 Jul;20(1-2):25–31. doi: 10.1007/BF00312761. [DOI] [PubMed] [Google Scholar]
  31. Resnick M. A., Kasimos J. N., Game J. C., Braun R. J., Roth R. M. Changes in DNA during meiosis in a repair-deficient mutant (rad 52) of yeast. Science. 1981 May 1;212(4494):543–545. doi: 10.1126/science.7010606. [DOI] [PubMed] [Google Scholar]
  32. Resnick M. A. The repair of double-strand breaks in DNA; a model involving recombination. J Theor Biol. 1976 Jun;59(1):97–106. doi: 10.1016/s0022-5193(76)80025-2. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Schultes N. P., Szostak J. W. Decreasing gradients of gene conversion on both sides of the initiation site for meiotic recombination at the ARG4 locus in yeast. Genetics. 1990 Dec;126(4):813–822. doi: 10.1093/genetics/126.4.813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Smithies O., Powers P. A. Gene conversions and their relation to homologous chromosome pairing. Philos Trans R Soc Lond B Biol Sci. 1986 Jan 29;312(1154):291–302. doi: 10.1098/rstb.1986.0008. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. 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]
  38. 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]
  39. Symington L. S., Brown A., Oliver S. G., Greenwell P., Petes T. D. Genetic analysis of a meiotic recombination hotspot on chromosome III of Saccharomyces cerevisiae. Genetics. 1991 Aug;128(4):717–727. doi: 10.1093/genetics/128.4.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. 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]
  41. 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]
  42. Yoshikawa A., Isono K. Chromosome III of Saccharomyces cerevisiae: an ordered clone bank, a detailed restriction map and analysis of transcripts suggest the presence of 160 genes. Yeast. 1990 Sep-Oct;6(5):383–401. doi: 10.1002/yea.320060504. [DOI] [PubMed] [Google Scholar]

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