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. 1999 Mar 15;18(6):1701–1711. doi: 10.1093/emboj/18.6.1701

The essential role of yeast topoisomerase III in meiosis depends on recombination.

S Gangloff 1, B de Massy 1, L Arthur 1, R Rothstein 1, F Fabre 1
PMCID: PMC1171256  PMID: 10075939

Abstract

Yeast cells mutant for TOP3, the gene encoding the evolutionary conserved type I-5' topoisomerase, display a wide range of phenotypes including altered cell cycle, hyper-recombination, abnormal gene expression, poor mating, chromosome instability and absence of sporulation. In this report, an analysis of the role of TOP3 in the meiotic process indicates that top3Delta mutants enter meiosis and complete the initial steps of recombination. However, reductional division does not occur. Deletion of the SPO11 gene, which prevents recombination between homologous chromosomes in meiosis I division, allows top3Delta mutants to form viable spores, indicating that Top3 is required to complete recombination successfully. A topoisomerase activity is involved in this process, since expression of bacterial TopA in yeast top3Delta mutants permits sporulation. The meiotic block is also partially suppressed by a deletion of SGS1, a gene encoding a helicase that interacts with Top3. We propose an essential role for Top3 in the processing of molecules generated during meiotic recombination.

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Selected References

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  1. Adachi Y., Luke M., Laemmli U. K. Chromosome assembly in vitro: topoisomerase II is required for condensation. Cell. 1991 Jan 11;64(1):137–148. doi: 10.1016/0092-8674(91)90215-k. [DOI] [PubMed] [Google Scholar]
  2. Bailis A. M., Arthur L., Rothstein R. Genome rearrangement in top3 mutants of Saccharomyces cerevisiae requires a functional RAD1 excision repair gene. Mol Cell Biol. 1992 Nov;12(11):4988–4993. doi: 10.1128/mcb.12.11.4988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baudat F., Nicolas A. Clustering of meiotic double-strand breaks on yeast chromosome III. Proc Natl Acad Sci U S A. 1997 May 13;94(10):5213–5218. doi: 10.1073/pnas.94.10.5213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bergerat A., de Massy B., Gadelle D., Varoutas P. C., Nicolas A., Forterre P. An atypical topoisomerase II from Archaea with implications for meiotic recombination. Nature. 1997 Mar 27;386(6623):414–417. doi: 10.1038/386414a0. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Castaño I. B., Brzoska P. M., Sadoff B. U., Chen H., Christman M. F. Mitotic chromosome condensation in the rDNA requires TRF4 and DNA topoisomerase I in Saccharomyces cerevisiae. Genes Dev. 1996 Oct 15;10(20):2564–2576. doi: 10.1101/gad.10.20.2564. [DOI] [PubMed] [Google Scholar]
  8. Champoux J. J. Mechanism of catalysis by eukaryotic DNA topoisomerase I. Adv Pharmacol. 1994;29A:71–82. doi: 10.1016/s1054-3589(08)60540-2. [DOI] [PubMed] [Google Scholar]
  9. Collins I., Newlon C. S. Chromosomal DNA replication initiates at the same origins in meiosis and mitosis. Mol Cell Biol. 1994 May;14(5):3524–3534. doi: 10.1128/mcb.14.5.3524. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. De Massy B., Baudat F., Nicolas A. Initiation of recombination in Saccharomyces cerevisiae haploid meiosis. Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):11929–11933. doi: 10.1073/pnas.91.25.11929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. DiNardo S., Voelkel K., Sternglanz R. DNA topoisomerase II mutant of Saccharomyces cerevisiae: topoisomerase II is required for segregation of daughter molecules at the termination of DNA replication. Proc Natl Acad Sci U S A. 1984 May;81(9):2616–2620. doi: 10.1073/pnas.81.9.2616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Downes C. S., Clarke D. J., Mullinger A. M., Giménez-Abián J. F., Creighton A. M., Johnson R. T. A topoisomerase II-dependent G2 cycle checkpoint in mammalian cells/. Nature. 1994 Dec 1;372(6505):467–470. doi: 10.1038/372467a0. [DOI] [PubMed] [Google Scholar]
  13. Drlica K. Bacterial topoisomerases and the control of DNA supercoiling. Trends Genet. 1990 Dec;6(12):433–437. doi: 10.1016/0168-9525(90)90306-q. [DOI] [PubMed] [Google Scholar]
  14. Duguet M. When helicase and topoisomerase meet! J Cell Sci. 1997 Jun;110(Pt 12):1345–1350. doi: 10.1242/jcs.110.12.1345. [DOI] [PubMed] [Google Scholar]
  15. Ellis N. A., Groden J., Ye T. Z., Straughen J., Lennon D. J., Ciocci S., Proytcheva M., German J. The Bloom's syndrome gene product is homologous to RecQ helicases. Cell. 1995 Nov 17;83(4):655–666. doi: 10.1016/0092-8674(95)90105-1. [DOI] [PubMed] [Google Scholar]
  16. Ezekiel U. R., Towler E. M., Wallis J. W., Zassenhaus H. P. Evidence for a nucleotide-dependent topoisomerase activity from yeast mitochondria. Curr Genet. 1994 Dec;27(1):31–37. doi: 10.1007/BF00326576. [DOI] [PubMed] [Google Scholar]
  17. Fabre F., Boulet A., Roman H. Gene conversion at different points in the mitotic cycle of Saccharomyces cerevisiae. Mol Gen Genet. 1984;195(1-2):139–143. doi: 10.1007/BF00332736. [DOI] [PubMed] [Google Scholar]
  18. Fritz E., Elsea S. H., Patel P. I., Meyn M. S. Overexpression of a truncated human topoisomerase III partially corrects multiple aspects of the ataxia-telangiectasia phenotype. Proc Natl Acad Sci U S A. 1997 Apr 29;94(9):4538–4542. doi: 10.1073/pnas.94.9.4538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gangloff S., Lieber M. R., Rothstein R. Transcription, topoisomerases and recombination. Experientia. 1994 Mar 15;50(3):261–269. doi: 10.1007/BF01924009. [DOI] [PubMed] [Google Scholar]
  20. Gangloff S., McDonald J. P., Bendixen C., Arthur L., Rothstein R. The yeast type I topoisomerase Top3 interacts with Sgs1, a DNA helicase homolog: a potential eukaryotic reverse gyrase. Mol Cell Biol. 1994 Dec;14(12):8391–8398. doi: 10.1128/mcb.14.12.8391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gangloff S., Zou H., Rothstein R. Gene conversion plays the major role in controlling the stability of large tandem repeats in yeast. EMBO J. 1996 Apr 1;15(7):1715–1725. [PMC free article] [PubMed] [Google Scholar]
  22. Giaever G. N., Wang J. C. Supercoiling of intracellular DNA can occur in eukaryotic cells. Cell. 1988 Dec 2;55(5):849–856. doi: 10.1016/0092-8674(88)90140-7. [DOI] [PubMed] [Google Scholar]
  23. Gilbertson L. A., Stahl F. W. A test of the double-strand break repair model for meiotic recombination in Saccharomyces cerevisiae. Genetics. 1996 Sep;144(1):27–41. doi: 10.1093/genetics/144.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Goto T., Wang J. C. Cloning of yeast TOP1, the gene encoding DNA topoisomerase I, and construction of mutants defective in both DNA topoisomerase I and DNA topoisomerase II. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7178–7182. doi: 10.1073/pnas.82.21.7178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Goto T., Wang J. C. Yeast DNA topoisomerase II is encoded by a single-copy, essential gene. Cell. 1984 Apr;36(4):1073–1080. doi: 10.1016/0092-8674(84)90057-6. [DOI] [PubMed] [Google Scholar]
  26. Hanai R., Caron P. R., Wang J. C. Human TOP3: a single-copy gene encoding DNA topoisomerase III. Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3653–3657. doi: 10.1073/pnas.93.8.3653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Holm C., Goto T., Wang J. C., Botstein D. DNA topoisomerase II is required at the time of mitosis in yeast. Cell. 1985 Jun;41(2):553–563. doi: 10.1016/s0092-8674(85)80028-3. [DOI] [PubMed] [Google Scholar]
  28. Holm C., Stearns T., Botstein D. DNA topoisomerase II must act at mitosis to prevent nondisjunction and chromosome breakage. Mol Cell Biol. 1989 Jan;9(1):159–168. doi: 10.1128/mcb.9.1.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Hsieh T., Lee M. P., Brown S. D. Structure of eukaryotic type I DNA topoisomerase. Adv Pharmacol. 1994;29A:191–200. doi: 10.1016/s1054-3589(08)60546-3. [DOI] [PubMed] [Google Scholar]
  30. Kadyk L. C., Hartwell L. H. Sister chromatids are preferred over homologs as substrates for recombinational repair in Saccharomyces cerevisiae. Genetics. 1992 Oct;132(2):387–402. doi: 10.1093/genetics/132.2.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kawasaki I., Bae Y. S., Eki T., Kim Y., Ikeda H. Homologous recombination of monkey alpha-satellite repeats in an in vitro simian virus 40 replication system: possible association of recombination with DNA replication. Mol Cell Biol. 1994 Jun;14(6):4173–4182. doi: 10.1128/mcb.14.6.4173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Kawasaki K., Minoshima S., Nakato E., Shibuya K., Shintani A., Schmeits J. L., Wang J., Shimizu N. One-megabase sequence analysis of the human immunoglobulin lambda gene locus. Genome Res. 1997 Mar;7(3):250–261. doi: 10.1101/gr.7.3.250. [DOI] [PubMed] [Google Scholar]
  33. Keeney S., Giroux C. N., Kleckner N. Meiosis-specific DNA double-strand breaks are catalyzed by Spo11, a member of a widely conserved protein family. Cell. 1997 Feb 7;88(3):375–384. doi: 10.1016/s0092-8674(00)81876-0. [DOI] [PubMed] [Google Scholar]
  34. Kim R. A., Caron P. R., Wang J. C. Effects of yeast DNA topoisomerase III on telomere structure. Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2667–2671. doi: 10.1073/pnas.92.7.2667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Kim R. A., Wang J. C. A subthreshold level of DNA topoisomerases leads to the excision of yeast rDNA as extrachromosomal rings. Cell. 1989 Jun 16;57(6):975–985. doi: 10.1016/0092-8674(89)90336-x. [DOI] [PubMed] [Google Scholar]
  36. Kim R. A., Wang J. C. Identification of the yeast TOP3 gene product as a single strand-specific DNA topoisomerase. J Biol Chem. 1992 Aug 25;267(24):17178–17185. [PubMed] [Google Scholar]
  37. 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]
  38. 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]
  39. Li W., Wang J. C. Mammalian DNA topoisomerase IIIalpha is essential in early embryogenesis. Proc Natl Acad Sci U S A. 1998 Feb 3;95(3):1010–1013. doi: 10.1073/pnas.95.3.1010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Lima C. D., Mondragón A. Mechanism of type II DNA topoisomerases: a tale of two gates. Structure. 1994 Jun 15;2(6):559–560. doi: 10.1016/s0969-2126(00)00055-1. [DOI] [PubMed] [Google Scholar]
  41. Liu L. F., Wang J. C. Supercoiling of the DNA template during transcription. Proc Natl Acad Sci U S A. 1987 Oct;84(20):7024–7027. doi: 10.1073/pnas.84.20.7024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. 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]
  43. Lydall D., Weinert T. Yeast checkpoint genes in DNA damage processing: implications for repair and arrest. Science. 1995 Dec 1;270(5241):1488–1491. doi: 10.1126/science.270.5241.1488. [DOI] [PubMed] [Google Scholar]
  44. 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]
  45. Nakayama K., Irino N., Nakayama H. The recQ gene of Escherichia coli K12: molecular cloning and isolation of insertion mutants. Mol Gen Genet. 1985;200(2):266–271. doi: 10.1007/BF00425434. [DOI] [PubMed] [Google Scholar]
  46. Puranam K. L., Blackshear P. J. Cloning and characterization of RECQL, a potential human homologue of the Escherichia coli DNA helicase RecQ. J Biol Chem. 1994 Nov 25;269(47):29838–29845. [PubMed] [Google Scholar]
  47. Puranam K. L., Kennington E., Sait S. N., Shows T. B., Rochelle J. M., Seldin M. F., Blackshear P. J. Chromosomal localization of the gene encoding the human DNA helicase RECQL and its mouse homologue. Genomics. 1995 Apr 10;26(3):595–598. doi: 10.1016/0888-7543(95)80181-k. [DOI] [PubMed] [Google Scholar]
  48. Roca J. The mechanisms of DNA topoisomerases. Trends Biochem Sci. 1995 Apr;20(4):156–160. doi: 10.1016/s0968-0004(00)88993-8. [DOI] [PubMed] [Google Scholar]
  49. Roeder G. S. Meiotic chromosomes: it takes two to tango. Genes Dev. 1997 Oct 15;11(20):2600–2621. doi: 10.1101/gad.11.20.2600. [DOI] [PubMed] [Google Scholar]
  50. 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]
  51. Rothstein R., Gangloff S. Hyper-recombination and Bloom's syndrome: microbes again provide clues about cancer. Genome Res. 1995 Dec;5(5):421–426. doi: 10.1101/gr.5.5.421. [DOI] [PubMed] [Google Scholar]
  52. 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]
  53. 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]
  54. 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]
  55. Schwacha A., Kleckner N. Interhomolog bias during meiotic recombination: meiotic functions promote a highly differentiated interhomolog-only pathway. Cell. 1997 Sep 19;90(6):1123–1135. doi: 10.1016/s0092-8674(00)80378-5. [DOI] [PubMed] [Google Scholar]
  56. Seki T., Seki M., Katada T., Enomoto T. Isolation of a cDNA encoding mouse DNA topoisomerase III which is highly expressed at the mRNA level in the testis. Biochim Biophys Acta. 1998 Mar 9;1396(2):127–131. doi: 10.1016/s0167-4781(97)00192-9. [DOI] [PubMed] [Google Scholar]
  57. 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]
  58. Sia R. A., Mitchell A. P. Stimulation of later functions of the yeast meiotic protein kinase Ime2p by the IDS2 gene product. Mol Cell Biol. 1995 Oct;15(10):5279–5287. doi: 10.1128/mcb.15.10.5279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Stewart E., Chapman C. R., Al-Khodairy F., Carr A. M., Enoch T. rqh1+, a fission yeast gene related to the Bloom's and Werner's syndrome genes, is required for reversible S phase arrest. EMBO J. 1997 May 15;16(10):2682–2692. doi: 10.1093/emboj/16.10.2682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Thomas B. J., Rothstein R. Elevated recombination rates in transcriptionally active DNA. Cell. 1989 Feb 24;56(4):619–630. doi: 10.1016/0092-8674(89)90584-9. [DOI] [PubMed] [Google Scholar]
  61. Thrash C., Bankier A. T., Barrell B. G., Sternglanz R. Cloning, characterization, and sequence of the yeast DNA topoisomerase I gene. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4374–4378. doi: 10.1073/pnas.82.13.4374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Thrash C., Voelkel K., DiNardo S., Sternglanz R. Identification of Saccharomyces cerevisiae mutants deficient in DNA topoisomerase I activity. J Biol Chem. 1984 Feb 10;259(3):1375–1377. [PubMed] [Google Scholar]
  63. Tsao Y. P., Wu H. Y., Liu L. F. Transcription-driven supercoiling of DNA: direct biochemical evidence from in vitro studies. Cell. 1989 Jan 13;56(1):111–118. doi: 10.1016/0092-8674(89)90989-6. [DOI] [PubMed] [Google Scholar]
  64. Umezu K., Nakayama K., Nakayama H. Escherichia coli RecQ protein is a DNA helicase. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5363–5367. doi: 10.1073/pnas.87.14.5363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Wallis J. W., Chrebet G., Brodsky G., Rolfe M., Rothstein R. A hyper-recombination mutation in S. cerevisiae identifies a novel eukaryotic topoisomerase. Cell. 1989 Jul 28;58(2):409–419. doi: 10.1016/0092-8674(89)90855-6. [DOI] [PubMed] [Google Scholar]
  66. Wang J. C. DNA topoisomerases. Annu Rev Biochem. 1996;65:635–692. doi: 10.1146/annurev.bi.65.070196.003223. [DOI] [PubMed] [Google Scholar]
  67. Wang Z., Dröge P. Differential control of transcription-induced and overall DNA supercoiling by eukaryotic topoisomerases in vitro. EMBO J. 1996 Feb 1;15(3):581–589. [PMC free article] [PubMed] [Google Scholar]
  68. Watt P. M., Hickson I. D., Borts R. H., Louis E. J. SGS1, a homologue of the Bloom's and Werner's syndrome genes, is required for maintenance of genome stability in Saccharomyces cerevisiae. Genetics. 1996 Nov;144(3):935–945. doi: 10.1093/genetics/144.3.935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  69. Watt P. M., Hickson I. D. Structure and function of type II DNA topoisomerases. Biochem J. 1994 Nov 1;303(Pt 3):681–695. doi: 10.1042/bj3030681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  70. Watt P. M., Louis E. J., Borts R. H., Hickson I. D. Sgs1: a eukaryotic homolog of E. coli RecQ that interacts with topoisomerase II in vivo and is required for faithful chromosome segregation. Cell. 1995 Apr 21;81(2):253–260. doi: 10.1016/0092-8674(95)90335-6. [DOI] [PubMed] [Google Scholar]
  71. Wu H. Y., Shyy S. H., Wang J. C., Liu L. F. Transcription generates positively and negatively supercoiled domains in the template. Cell. 1988 May 6;53(3):433–440. doi: 10.1016/0092-8674(88)90163-8. [DOI] [PubMed] [Google Scholar]
  72. 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]
  73. Yan H., Chen C. Y., Kobayashi R., Newport J. Replication focus-forming activity 1 and the Werner syndrome gene product. Nat Genet. 1998 Aug;19(4):375–378. doi: 10.1038/1263. [DOI] [PubMed] [Google Scholar]
  74. Young C. S., Cachianes G., Munz P., Silverstein S. Replication and recombination in adenovirus-infected cells are temporally and functionally related. J Virol. 1984 Sep;51(3):571–577. doi: 10.1128/jvi.51.3.571-577.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  75. Yu C. E., Oshima J., Fu Y. H., Wijsman E. M., Hisama F., Alisch R., Matthews S., Nakura J., Miki T., Ouais S. Positional cloning of the Werner's syndrome gene. Science. 1996 Apr 12;272(5259):258–262. doi: 10.1126/science.272.5259.258. [DOI] [PubMed] [Google Scholar]
  76. Zou H., Rothstein R. Holliday junctions accumulate in replication mutants via a RecA homolog-independent mechanism. Cell. 1997 Jul 11;90(1):87–96. doi: 10.1016/s0092-8674(00)80316-5. [DOI] [PubMed] [Google Scholar]
  77. 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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