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. 1997 Jul;146(3):817–834. doi: 10.1093/genetics/146.3.817

Mutations in Saccharomyces Cerevisiae That Block Meiotic Prophase Chromosome Metabolism and Confer Cell Cycle Arrest at Pachytene Identify Two New Meiosis-Specific Genes Sae1 and Sae3

AHZ McKee 1, N Kleckner 1
PMCID: PMC1208053  PMID: 9215889

Abstract

Two new meiosis-specific genes, SAE1 and SAE3, have been identified in a screen for mutations that confer an intermediate block in meiotic prophase. Such mutations confer a block to spore formation that is circumvented by addition of a mutation that eliminates meiotic recombination initiation and other aspects of chromosome metabolism, i.e., spo11. We show that sae1-1 and sae3-1 mutations each confer a distinct defect in meiotic recombination. sae1-1 produces recombinants but very slowly and ultimately to less than half the wild-type level; sae3-1 makes persistent hyper-resected meiotic double-strand breaks and has a severe defect in formation of recombinants. Both mutants arrest at the pachytene stage of meiotic prophase, sae1-1 temporarily and sae3-1 permanently. The phenotypes conferred by sae3-1 are similar to those conferred by mutation of the yeast RecA homologue DMC1, suggesting that SAE3 and DMC1 act at the same step(s) of chromosome metabolism. These results provide further evidence that intermediate blocks to prophase chromosome metabolism cause cell-cycle arrest. SAE1 encodes a 208-residue protein homologous to vertebrate mRNA cap-binding protein 20. SAE3 corresponds to a meiosis-specific RNA encoding an unusually short open reading frame of 50 codons.

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Selected References

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  1. Alani E., Cao L., Kleckner N. A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains. Genetics. 1987 Aug;116(4):541–545. doi: 10.1534/genetics.112.541.test. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Becker D. M., Guarente L. High-efficiency transformation of yeast by electroporation. Methods Enzymol. 1991;194:182–187. doi: 10.1016/0076-6879(91)94015-5. [DOI] [PubMed] [Google Scholar]
  3. Betz R., Crabb J. W., Meyer H. E., Wittig R., Duntze W. Amino acid sequences of a-factor mating peptides from Saccharomyces cerevisiae. J Biol Chem. 1987 Jan 15;262(2):546–548. [PubMed] [Google Scholar]
  4. Bishop D. K., Park D., Xu L., Kleckner N. DMC1: a meiosis-specific yeast homolog of E. coli recA required for recombination, synaptonemal complex formation, and cell cycle progression. Cell. 1992 May 1;69(3):439–456. doi: 10.1016/0092-8674(92)90446-j. [DOI] [PubMed] [Google Scholar]
  5. Bishop D. K. RecA homologs Dmc1 and Rad51 interact to form multiple nuclear complexes prior to meiotic chromosome synapsis. Cell. 1994 Dec 16;79(6):1081–1092. doi: 10.1016/0092-8674(94)90038-8. [DOI] [PubMed] [Google Scholar]
  6. Borts R. H., Lichten M., Haber J. E. Analysis of meiosis-defective mutations in yeast by physical monitoring of recombination. Genetics. 1986 Jul;113(3):551–567. doi: 10.1093/genetics/113.3.551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Briza P., Breitenbach M., Ellinger A., Segall J. Isolation of two developmentally regulated genes involved in spore wall maturation in Saccharomyces cerevisiae. Genes Dev. 1990 Oct;4(10):1775–1789. doi: 10.1101/gad.4.10.1775. [DOI] [PubMed] [Google Scholar]
  8. Bullard S. A., Kim S., Galbraith A. M., Malone R. E. Double strand breaks at the HIS2 recombination hot spot in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):13054–13059. doi: 10.1073/pnas.93.23.13054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Burd C. G., Dreyfuss G. Conserved structures and diversity of functions of RNA-binding proteins. Science. 1994 Jul 29;265(5172):615–621. doi: 10.1126/science.8036511. [DOI] [PubMed] [Google Scholar]
  10. Bähler J., Hagens G., Holzinger G., Scherthan H., Heyer W. D. Saccharomyces cerevisiae cells lacking the homologous pairing protein p175SEP1 arrest at pachytene during meiotic prophase. Chromosoma. 1994 Apr;103(2):129–141. doi: 10.1007/BF00352322. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Chi J. H., Roos J., Dean N. The OST4 gene of Saccharomyces cerevisiae encodes an unusually small protein required for normal levels of oligosaccharyltransferase activity. J Biol Chem. 1996 Feb 9;271(6):3132–3140. doi: 10.1074/jbc.271.6.3132. [DOI] [PubMed] [Google Scholar]
  13. Engebrecht J. A., Voelkel-Meiman K., Roeder G. S. Meiosis-specific RNA splicing in yeast. Cell. 1991 Sep 20;66(6):1257–1268. doi: 10.1016/0092-8674(91)90047-3. [DOI] [PubMed] [Google Scholar]
  14. Engebrecht J., Roeder G. S. Yeast mer1 mutants display reduced levels of meiotic recombination. Genetics. 1989 Feb;121(2):237–247. doi: 10.1093/genetics/121.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Goldman A. S., Lichten M. The efficiency of meiotic recombination between dispersed sequences in Saccharomyces cerevisiae depends upon their chromosomal location. Genetics. 1996 Sep;144(1):43–55. doi: 10.1093/genetics/144.1.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Izaurralde E., Lewis J., Gamberi C., Jarmolowski A., McGuigan C., Mattaj I. W. A cap-binding protein complex mediating U snRNA export. Nature. 1995 Aug 24;376(6542):709–712. doi: 10.1038/376709a0. [DOI] [PubMed] [Google Scholar]
  17. Izaurralde E., Lewis J., McGuigan C., Jankowska M., Darzynkiewicz E., Mattaj I. W. A nuclear cap binding protein complex involved in pre-mRNA splicing. Cell. 1994 Aug 26;78(4):657–668. doi: 10.1016/0092-8674(94)90530-4. [DOI] [PubMed] [Google Scholar]
  18. Jarmolowski A., Boelens W. C., Izaurralde E., Mattaj I. W. Nuclear export of different classes of RNA is mediated by specific factors. J Cell Biol. 1994 Mar;124(5):627–635. doi: 10.1083/jcb.124.5.627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Johnston M., Andrews S., Brinkman R., Cooper J., Ding H., Dover J., Du Z., Favello A., Fulton L., Gattung S. Complete nucleotide sequence of Saccharomyces cerevisiae chromosome VIII. Science. 1994 Sep 30;265(5181):2077–2082. doi: 10.1126/science.8091229. [DOI] [PubMed] [Google Scholar]
  20. Kane S. M., Roth R. Carbohydrate metabolism during ascospore development in yeast. J Bacteriol. 1974 Apr;118(1):8–14. doi: 10.1128/jb.118.1.8-14.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kataoka N., Ohno M., Moda I., Shimura Y. Identification of the factors that interact with NCBP, an 80 kDa nuclear cap binding protein. Nucleic Acids Res. 1995 Sep 25;23(18):3638–3641. doi: 10.1093/nar/23.18.3638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Klapholz S., Waddell C. S., Esposito R. E. The role of the SPO11 gene in meiotic recombination in yeast. Genetics. 1985 Jun;110(2):187–216. doi: 10.1093/genetics/110.2.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kleckner N. Meiosis: how could it work? Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8167–8174. doi: 10.1073/pnas.93.16.8167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Kurjan J., Herskowitz I. Structure of a yeast pheromone gene (MF alpha): a putative alpha-factor precursor contains four tandem copies of mature alpha-factor. Cell. 1982 Oct;30(3):933–943. doi: 10.1016/0092-8674(82)90298-7. [DOI] [PubMed] [Google Scholar]
  26. Lichten M., Goldman A. S. Meiotic recombination hotspots. Annu Rev Genet. 1995;29:423–444. doi: 10.1146/annurev.ge.29.120195.002231. [DOI] [PubMed] [Google Scholar]
  27. Lydall D., Nikolsky Y., Bishop D. K., Weinert T. A meiotic recombination checkpoint controlled by mitotic checkpoint genes. Nature. 1996 Oct 31;383(6603):840–843. doi: 10.1038/383840a0. [DOI] [PubMed] [Google Scholar]
  28. Malone R. E., Esposito R. E. Recombinationless meiosis in Saccharomyces cerevisiae. Mol Cell Biol. 1981 Oct;1(10):891–901. doi: 10.1128/mcb.1.10.891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mattaj I. W. RNA recognition: a family matter? Cell. 1993 Jun 4;73(5):837–840. doi: 10.1016/0092-8674(93)90265-r. [DOI] [PubMed] [Google Scholar]
  30. McKee A. H., Kleckner N. A general method for identifying recessive diploid-specific mutations in Saccharomyces cerevisiae, its application to the isolation of mutants blocked at intermediate stages of meiotic prophase and characterization of a new gene SAE2. Genetics. 1997 Jul;146(3):797–816. doi: 10.1093/genetics/146.3.797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Michaelis S., Herskowitz I. The a-factor pheromone of Saccharomyces cerevisiae is essential for mating. Mol Cell Biol. 1988 Mar;8(3):1309–1318. doi: 10.1128/mcb.8.3.1309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Nakagawa T., Ogawa H. Involvement of the MRE2 gene of yeast in formation of meiosis-specific double-strand breaks and crossover recombination through RNA splicing. Genes Cells. 1997 Jan;2(1):65–79. doi: 10.1046/j.1365-2443.1997.d01-283.x. [DOI] [PubMed] [Google Scholar]
  33. Navarre C., Catty P., Leterme S., Dietrich F., Goffeau A. Two distinct genes encode small isoproteolipids affecting plasma membrane H(+)-ATPase activity of Saccharomyces cerevisiae. J Biol Chem. 1994 Aug 19;269(33):21262–21268. [PubMed] [Google Scholar]
  34. Navarre C., Ghislain M., Leterme S., Ferroud C., Dufour J. P., Goffeau A. Purification and complete sequence of a small proteolipid associated with the plasma membrane H(+)-ATPase of Saccharomyces cerevisiae. J Biol Chem. 1992 Mar 25;267(9):6425–6428. [PubMed] [Google Scholar]
  35. Nikawa J., Yamashita S. IRE1 encodes a putative protein kinase containing a membrane-spanning domain and is required for inositol phototrophy in Saccharomyces cerevisiae. Mol Microbiol. 1992 Jun;6(11):1441–1446. doi: 10.1111/j.1365-2958.1992.tb00864.x. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Prinz S., Amon A., Klein F. Isolation of COM1, a new gene required to complete meiotic double-strand break-induced recombination in Saccharomyces cerevisiae. Genetics. 1997 Jul;146(3):781–795. doi: 10.1093/genetics/146.3.781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Resnick M. A., Nitiss J., Edwards C., Malone R. E. Meiosis can induce recombination in rad52 mutants of Saccharomyces cerevisiae. Genetics. 1986 Jul;113(3):531–550. doi: 10.1093/genetics/113.3.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Rocco V., Nicolas A. Sensing of DNA non-homology lowers the initiation of meiotic recombination in yeast. Genes Cells. 1996 Jul;1(7):645–661. doi: 10.1046/j.1365-2443.1996.00256.x. [DOI] [PubMed] [Google Scholar]
  40. Rockmill B., Sym M., Scherthan H., Roeder G. S. Roles for two RecA homologs in promoting meiotic chromosome synapsis. Genes Dev. 1995 Nov 1;9(21):2684–2695. doi: 10.1101/gad.9.21.2684. [DOI] [PubMed] [Google Scholar]
  41. 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]
  42. Roeder G. S. Sex and the single cell: meiosis in yeast. Proc Natl Acad Sci U S A. 1995 Nov 7;92(23):10450–10456. doi: 10.1073/pnas.92.23.10450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Rose D., Holm C. Meiosis-specific arrest revealed in DNA topoisomerase II mutants. Mol Cell Biol. 1993 Jun;13(6):3445–3455. doi: 10.1128/mcb.13.6.3445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Rose D., Thomas W., Holm C. Segregation of recombined chromosomes in meiosis I requires DNA topoisomerase II. Cell. 1990 Mar 23;60(6):1009–1017. doi: 10.1016/0092-8674(90)90349-j. [DOI] [PubMed] [Google Scholar]
  45. Rose M. D., Novick P., Thomas J. H., Botstein D., Fink G. R. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector. Gene. 1987;60(2-3):237–243. doi: 10.1016/0378-1119(87)90232-0. [DOI] [PubMed] [Google Scholar]
  46. SHERMAN F., ROMAN H. Evidence for two types of allelic recombination in yeast. Genetics. 1963 Feb;48:255–261. doi: 10.1093/genetics/48.2.255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. 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]
  48. 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]
  49. Shinohara A., Ogawa H., Ogawa T. Rad51 protein involved in repair and recombination in S. cerevisiae is a RecA-like protein. Cell. 1992 May 1;69(3):457–470. doi: 10.1016/0092-8674(92)90447-k. [DOI] [PubMed] [Google Scholar]
  50. Shuster E. O., Byers B. Pachytene arrest and other meiotic effects of the start mutations in Saccharomyces cerevisiae. Genetics. 1989 Sep;123(1):29–43. doi: 10.1093/genetics/123.1.29. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Singh A., Chen E. Y., Lugovoy J. M., Chang C. N., Hitzeman R. A., Seeburg P. H. Saccharomyces cerevisiae contains two discrete genes coding for the alpha-factor pheromone. Nucleic Acids Res. 1983 Jun 25;11(12):4049–4063. doi: 10.1093/nar/11.12.4049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Storlazzi A., Xu L., Schwacha A., Kleckner N. Synaptonemal complex (SC) component Zip1 plays a role in meiotic recombination independent of SC polymerization along the chromosomes. Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):9043–9048. doi: 10.1073/pnas.93.17.9043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Strathern J. N., Herskowitz I. Asymmetry and directionality in production of new cell types during clonal growth: the switching pattern of homothallic yeast. Cell. 1979 Jun;17(2):371–381. doi: 10.1016/0092-8674(79)90163-6. [DOI] [PubMed] [Google Scholar]
  55. Sym M., Engebrecht J. A., Roeder G. S. ZIP1 is a synaptonemal complex protein required for meiotic chromosome synapsis. Cell. 1993 Feb 12;72(3):365–378. doi: 10.1016/0092-8674(93)90114-6. [DOI] [PubMed] [Google Scholar]
  56. Tanaka T., Kita H., Murakami T., Narita K. Purification and amino acid sequence of mating factor from Saccharomyces cerevisiae. J Biochem. 1977 Dec;82(6):1681–1687. doi: 10.1093/oxfordjournals.jbchem.a131864. [DOI] [PubMed] [Google Scholar]
  57. Terasawa M., Shinohara A., Hotta Y., Ogawa H., Ogawa T. Localization of RecA-like recombination proteins on chromosomes of the lily at various meiotic stages. Genes Dev. 1995 Apr 15;9(8):925–934. doi: 10.1101/gad.9.8.925. [DOI] [PubMed] [Google Scholar]
  58. Tishkoff D. X., Rockmill B., Roeder G. S., Kolodner R. D. The sep1 mutant of Saccharomyces cerevisiae arrests in pachytene and is deficient in meiotic recombination. Genetics. 1995 Feb;139(2):495–509. doi: 10.1093/genetics/139.2.495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Vincent A., Newnam G., Liebman S. W. The yeast translational allosuppressor, SAL6: a new member of the PP1-like phosphatase family with a long serine-rich N-terminal extension. Genetics. 1994 Nov;138(3):597–608. doi: 10.1093/genetics/138.3.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Watanabe Y., Yamamoto M. S. pombe mei2+ encodes an RNA-binding protein essential for premeiotic DNA synthesis and meiosis I, which cooperates with a novel RNA species meiRNA. Cell. 1994 Aug 12;78(3):487–498. doi: 10.1016/0092-8674(94)90426-x. [DOI] [PubMed] [Google Scholar]
  61. Weiner B. M., Kleckner N. Chromosome pairing via multiple interstitial interactions before and during meiosis in yeast. Cell. 1994 Jul 1;77(7):977–991. doi: 10.1016/0092-8674(94)90438-3. [DOI] [PubMed] [Google Scholar]
  62. Xu L., Ajimura M., Padmore R., Klein C., Kleckner N. NDT80, a meiosis-specific gene required for exit from pachytene in Saccharomyces cerevisiae. Mol Cell Biol. 1995 Dec;15(12):6572–6581. doi: 10.1128/mcb.15.12.6572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Xu L., Kleckner N. Sequence non-specific double-strand breaks and interhomolog interactions prior to double-strand break formation at a meiotic recombination hot spot in yeast. EMBO J. 1995 Oct 16;14(20):5115–5128. doi: 10.1002/j.1460-2075.1995.tb00194.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Xu L., Weiner B. M., Kleckner N. Meiotic cells monitor the status of the interhomolog recombination complex. Genes Dev. 1997 Jan 1;11(1):106–118. doi: 10.1101/gad.11.1.106. [DOI] [PubMed] [Google Scholar]
  65. Yoon H., Donahue T. F. Control of translation initiation in Saccharomyces cerevisiae. Mol Microbiol. 1992 Jun;6(11):1413–1419. doi: 10.1111/j.1365-2958.1992.tb00861.x. [DOI] [PubMed] [Google Scholar]
  66. von Wettstein D., Rasmussen S. W., Holm P. B. The synaptonemal complex in genetic segregation. Annu Rev Genet. 1984;18:331–413. doi: 10.1146/annurev.ge.18.120184.001555. [DOI] [PubMed] [Google Scholar]

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