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. 1994 Nov 1;13(21):5212–5219. doi: 10.1002/j.1460-2075.1994.tb06852.x

Preferential strand transfer and hybrid DNA formation at the recombination hotspot ade6-M26 of Schizosaccharomyces pombe.

P Schär 1, J Kohli 1
PMCID: PMC395471  PMID: 7957086

Abstract

The ade6-M26 mutation of Schizosaccharomyces pombe stimulates intragenic and intergenic meiotic recombination. M26 is a single base pair change creating a specific heptanucleotide sequence that is crucial for recombination hotspot activity. This sequence is recognized by proteins that may facilitate rate-limiting steps of recombination at the ade6 locus. To start the elucidation of the intermediate DNA structures formed during M26 recombination, we have analyzed the aberrant segregation patterns of two G to C transversion mutations flanking the heptanucleotide sequence in crosses homozygous for M26. At both sites the level of post-meiotic segregation is typical for G to C transversion mutations in S. pombe in general. Quantitative treatment of the data provides strong evidence for heteroduplex DNA being the major recombination intermediate at the M26 site. We can now exclude a double-strand gap repair mechanism to account for gene conversion across the recombination hotspot. Furthermore, the vast majority (> 95%) of the heteroduplexes covering either of the G to C transversion sites are produced by transfer of the transcribed DNA strand. These results are consistent with ade6-M26 creating an initiation site for gene conversion by the introduction of a single-strand or a double-strand break in its vicinity, followed by transfer of the transcribed DNA strands for heteroduplex DNA formation.

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Selected References

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

  1. Bishop D. K., Andersen J., Kolodner R. D. Specificity of mismatch repair following transformation of Saccharomyces cerevisiae with heteroduplex plasmid DNA. Proc Natl Acad Sci U S A. 1989 May;86(10):3713–3717. doi: 10.1073/pnas.86.10.3713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bähler J., Schuchert P., Grimm C., Kohli J. Synchronized meiosis and recombination in fission yeast: observations with pat1-114 diploid cells. Curr Genet. 1991 Jun;19(6):445–451. doi: 10.1007/BF00312735. [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. 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]
  5. 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]
  6. Fogel S., Mortimer R., Lusnak K., Tavares F. Meiotic gene conversion: a signal of the basic recombination event in yeast. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1325–1341. doi: 10.1101/sqb.1979.043.01.152. [DOI] [PubMed] [Google Scholar]
  7. Goldman S. L., Smallets S. Site specific induction of gene conversion: the effects of homozygosity of the ade6 mutant M26 of Schizosaccharomyces pombe on meiotic gene conversion. Mol Gen Genet. 1979 Jun 20;173(3):221–225. doi: 10.1007/BF00268632. [DOI] [PubMed] [Google Scholar]
  8. Grimm C., Bähler J., Kohli J. M26 recombinational hotspot and physical conversion tract analysis in the ade6 gene of Schizosaccharomyces pombe. Genetics. 1994 Jan;136(1):41–51. doi: 10.1093/genetics/136.1.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gutz H. Site Specific Induction of Gene Conversion in SCHIZOSACCHAROMYCES POMBE. Genetics. 1971 Nov;69(3):317–337. doi: 10.1093/genetics/69.3.317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kramer B., Kramer W., Williamson M. S., Fogel S. Heteroduplex DNA correction in Saccharomyces cerevisiae is mismatch specific and requires functional PMS genes. Mol Cell Biol. 1989 Oct;9(10):4432–4440. doi: 10.1128/mcb.9.10.4432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lichten M., Goyon C., Schultes N. P., Treco D., Szostak J. W., Haber J. E., Nicolas A. Detection of heteroduplex DNA molecules among the products of Saccharomyces cerevisiae meiosis. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7653–7657. doi: 10.1073/pnas.87.19.7653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. 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]
  15. 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]
  16. Nicolas A., Petes T. D. Polarity of meiotic gene conversion in fungi: contrasting views. Experientia. 1994 Mar 15;50(3):242–252. doi: 10.1007/BF01924007. [DOI] [PubMed] [Google Scholar]
  17. Nicolas A., Treco D., Schultes N. P., Szostak J. W. An initiation site for meiotic gene conversion in the yeast Saccharomyces cerevisiae. Nature. 1989 Mar 2;338(6210):35–39. doi: 10.1038/338035a0. [DOI] [PubMed] [Google Scholar]
  18. Ponticelli A. S., Sena E. P., Smith G. R. Genetic and physical analysis of the M26 recombination hotspot of Schizosaccharomyces pombe. Genetics. 1988 Jul;119(3):491–497. doi: 10.1093/genetics/119.3.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Reenan R. A., Kolodner R. D. Characterization of insertion mutations in the Saccharomyces cerevisiae MSH1 and MSH2 genes: evidence for separate mitochondrial and nuclear functions. Genetics. 1992 Dec;132(4):975–985. doi: 10.1093/genetics/132.4.975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schuchert P., Langsford M., Käslin E., Kohli J. A specific DNA sequence is required for high frequency of recombination in the ade6 gene of fission yeast. EMBO J. 1991 Aug;10(8):2157–2163. doi: 10.1002/j.1460-2075.1991.tb07750.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schär P., Kohli J. Marker effects of G to C transversions on intragenic recombination and mismatch repair in Schizosaccharomyces pombe. Genetics. 1993 Apr;133(4):825–835. doi: 10.1093/genetics/133.4.825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schär P., Munz P., Kohli J. Meiotic mismatch repair quantified on the basis of segregation patterns in Schizosaccharomyces pombe. Genetics. 1993 Apr;133(4):815–824. doi: 10.1093/genetics/133.4.815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. Szankasi P., Heyer W. D., Schuchert P., Kohli J. DNA sequence analysis of the ade6 gene of Schizosaccharomyces pombe. Wild-type and mutant alleles including the recombination host spot allele ade6-M26. J Mol Biol. 1988 Dec 20;204(4):917–925. doi: 10.1016/0022-2836(88)90051-4. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Wahls W. P., Smith G. R. A heteromeric protein that binds to a meiotic homologous recombination hot spot: correlation of binding and hot spot activity. Genes Dev. 1994 Jul 15;8(14):1693–1702. doi: 10.1101/gad.8.14.1693. [DOI] [PubMed] [Google Scholar]
  29. White J. H., Lusnak K., Fogel S. Mismatch-specific post-meiotic segregation frequency in yeast suggests a heteroduplex recombination intermediate. Nature. 1985 May 23;315(6017):350–352. doi: 10.1038/315350a0. [DOI] [PubMed] [Google Scholar]
  30. Williamson M. S., Game J. C., Fogel S. Meiotic gene conversion mutants in Saccharomyces cerevisiae. I. Isolation and characterization of pms1-1 and pms1-2. Genetics. 1985 Aug;110(4):609–646. doi: 10.1093/genetics/110.4.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. de Massy B., Nicolas A. The control in cis of the position and the amount of the ARG4 meiotic double-strand break of Saccharomyces cerevisiae. EMBO J. 1993 Apr;12(4):1459–1466. doi: 10.1002/j.1460-2075.1993.tb05789.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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