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. 1994 Jan;136(1):41–51. doi: 10.1093/genetics/136.1.41

M26 Recombinational Hotspot and Physical Conversion Tract Analysis in the Ade6 Gene of Schizosaccharomyces Pombe

C Grimm 1, J Bahler 1, J Kohli 1
PMCID: PMC1205790  PMID: 7908005

Abstract

At the ade6 locus of Schizosaccharomyces pombe flanking markers have been introduced as well as five silent restriction site polymorphisms: four in the 5' upstream region and one in the middle of the gene. The mutations ade6-706, ade6-M26 (both at the 5' end) and ade6-51 (middle of the gene) were used as partners for crosses with the 3' mutation ade6-469. From these three types of crosses, wild-type recombinants were selected and analyzed genetically to assess association with crossing-over and physically to determine conversion tract lengths. The introduced restriction site polymorphisms (five vs. only one) neither influenced the pattern of recombinant types nor the distribution of conversion tracts. The hotspot mutation M26 enhances crossing-over and conversion to the same proportion. M26 not only stimulates conversion at the 5' end, but does this also (to a lower extent) at the 3' end of ade6 at a distance of more than 1 kb. The majority of meiotic conversion tracts are continuous and postmeiotic segregation of polymorphic sites is rare. Conversion tracts are slightly shorter with M26 in comparison with its control 706. The mean minimal length of tracts varies from 670 bp (M26) to 890 bp (706) to 1290 bp (51). It is concluded that M26 acts as an initiation site of recombination or enhances initiation of recombination. M26 does not act by termination of conversion. A region of recombination initiation exists at the 5' end of the ade6 gene also in the absence of the ade6-M26 hotspot mutation.

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Selected References

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  1. Ahn B. Y., Livingston D. M. Mitotic gene conversion lengths, coconversion patterns, and the incidence of reciprocal recombination in a Saccharomyces cerevisiae plasmid system. Mol Cell Biol. 1986 Nov;6(11):3685–3693. doi: 10.1128/mcb.6.11.3685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Borts R. H., Haber J. E. Length and distribution of meiotic gene conversion tracts and crossovers in Saccharomyces cerevisiae. Genetics. 1989 Sep;123(1):69–80. doi: 10.1093/genetics/123.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Borts R. H., Haber J. E. Meiotic recombination in yeast: alteration by multiple heterozygosities. Science. 1987 Sep 18;237(4821):1459–1465. doi: 10.1126/science.2820060. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Cummins J. E., Mitchison J. M. Adenine uptake and pool formation in the fission yeast Schizosaccharomyces pombe. Biochim Biophys Acta. 1967 Feb 7;136(1):108–120. doi: 10.1016/0304-4165(67)90326-1. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Fan J. B., Grothues D., Smith C. L. Alignment of Sfi I sites with the Not I restriction map of Schizosaccharomyces pombe genome. Nucleic Acids Res. 1991 Nov 25;19(22):6289–6294. doi: 10.1093/nar/19.22.6289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Grimm C., Kohli J., Murray J., Maundrell K. Genetic engineering of Schizosaccharomyces pombe: a system for gene disruption and replacement using the ura4 gene as a selectable marker. Mol Gen Genet. 1988 Dec;215(1):81–86. doi: 10.1007/BF00331307. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Keil R. L., Roeder G. S. Cis-acting, recombination-stimulating activity in a fragment of the ribosomal DNA of S. cerevisiae. Cell. 1984 Dec;39(2 Pt 1):377–386. doi: 10.1016/0092-8674(84)90016-3. [DOI] [PubMed] [Google Scholar]
  12. Malone R. E., Bullard S., Lundquist S., Kim S., Tarkowski T. A meiotic gene conversion gradient opposite to the direction of transcription. Nature. 1992 Sep 10;359(6391):154–155. doi: 10.1038/359154a0. [DOI] [PubMed] [Google Scholar]
  13. Munz P., Leupold U. Gene conversion in nonsense suppressors of Schizosaccharomyces pombe. I. The influence of the genetic background and of three mutant genes (rad2, mut1 and mut2) on the frequency of the post-meiotic segregation. Mol Gen Genet. 1979 Feb 26;170(2):145–148. doi: 10.1007/BF00337789. [DOI] [PubMed] [Google Scholar]
  14. Nakaseko Y., Adachi Y., Funahashi S., Niwa O., Yanagida M. Chromosome walking shows a highly homologous repetitive sequence present in all the centromere regions of fission yeast. EMBO J. 1986 May;5(5):1011–1021. doi: 10.1002/j.1460-2075.1986.tb04316.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Petes T. D., Botstein D. Simple Mendelian inheritance of the reiterated ribosomal DNA of yeast. Proc Natl Acad Sci U S A. 1977 Nov;74(11):5091–5095. doi: 10.1073/pnas.74.11.5091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ponticelli A. S., Smith G. R. Chromosomal context dependence of a eukaryotic recombinational hot spot. Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):227–231. doi: 10.1073/pnas.89.1.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ponticelli A. S., Smith G. R. Meiotic recombination-deficient mutants of Schizosaccharomyces pombe. Genetics. 1989 Sep;123(1):45–54. doi: 10.1093/genetics/123.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schuchert P., Kohli J. The Ade6-M26 Mutation of Schizosaccharomyces Pombe Increases the Frequency of Crossing over. Genetics. 1988 Jul;119(3):507–515. doi: 10.1093/genetics/119.3.507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schultes N. P., Szostak J. W. A poly(dA.dT) tract is a component of the recombination initiation site at the ARG4 locus in Saccharomyces cerevisiae. Mol Cell Biol. 1991 Jan;11(1):322–328. doi: 10.1128/mcb.11.1.322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  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. Stewart S. E., Roeder G. S. Transcription by RNA polymerase I stimulates mitotic recombination in Saccharomyces cerevisiae. Mol Cell Biol. 1989 Aug;9(8):3464–3472. doi: 10.1128/mcb.9.8.3464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. 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]
  28. 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]
  29. 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]
  30. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  31. White M. A., Wierdl M., Detloff P., Petes T. D. DNA-binding protein RAP1 stimulates meiotic recombination at the HIS4 locus in yeast. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9755–9759. doi: 10.1073/pnas.88.21.9755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wright A. P., Maundrell K., Shall S. Transformation of Schizosaccharomyces pombe by non-homologous, unstable integration of plasmids in the genome. Curr Genet. 1986;10(7):503–508. doi: 10.1007/BF00447383. [DOI] [PubMed] [Google Scholar]

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