Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1994 Mar;14(3):1796–1805. doi: 10.1128/mcb.14.3.1796

Specific initiation at an origin of replication from Schizosaccharomyces pombe.

M S Caddle 1, M P Calos 1
PMCID: PMC358537  PMID: 8114712

Abstract

Using a genetic assay for efficient autonomous replication, we have isolated from Schizosaccharomyces pombe a 6.2-kb fragment which shows the properties expected of an origin of DNA replication in S. pombe. A 2.8-kb subclone of the fragment has the same replication properties. Two-dimensional gel analysis of replication intermediates throughout plasmids carrying the 6.2- or 2.8-kb fragments shows that replication initiates only in a specific region, which can be localized to within several hundred base pairs, in the fragments. This region is also a site of replication initiation in the S. pombe chromosome where the fragments normally reside. These results provide strong evidence that initiation of replication in S. pombe is localized and mediated by specific DNA sequence signals.

Full text

PDF
1796

Images in this article

1798Image
on p.1798
1799Image
on p.1799
1801Image
on p.1801
1802Image
on p.1802
1802Image
on p.1802

Selected References

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

  1. Bell S. P., Stillman B. ATP-dependent recognition of eukaryotic origins of DNA replication by a multiprotein complex. Nature. 1992 May 14;357(6374):128–134. doi: 10.1038/357128a0. [DOI] [PubMed] [Google Scholar]
  2. Brewer B. J., Fangman W. L. The localization of replication origins on ARS plasmids in S. cerevisiae. Cell. 1987 Nov 6;51(3):463–471. doi: 10.1016/0092-8674(87)90642-8. [DOI] [PubMed] [Google Scholar]
  3. Brewer B. J., Lockshon D., Fangman W. L. The arrest of replication forks in the rDNA of yeast occurs independently of transcription. Cell. 1992 Oct 16;71(2):267–276. doi: 10.1016/0092-8674(92)90355-g. [DOI] [PubMed] [Google Scholar]
  4. Caddle M. S., Calos M. P. Analysis of the autonomous replication behavior in human cells of the dihydrofolate reductase putative chromosomal origin of replication. Nucleic Acids Res. 1992 Nov 25;20(22):5971–5978. doi: 10.1093/nar/20.22.5971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Celniker S. E., Sweder K., Srienc F., Bailey J. E., Campbell J. L. Deletion mutations affecting autonomously replicating sequence ARS1 of Saccharomyces cerevisiae. Mol Cell Biol. 1984 Nov;4(11):2455–2466. doi: 10.1128/mcb.4.11.2455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clarke L., Amstutz H., Fishel B., Carbon J. Analysis of centromeric DNA in the fission yeast Schizosaccharomyces pombe. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8253–8257. doi: 10.1073/pnas.83.21.8253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dijkwel P. A., Hamlin J. L. Initiation of DNA replication in the dihydrofolate reductase locus is confined to the early S period in CHO cells synchronized with the plant amino acid mimosine. Mol Cell Biol. 1992 Sep;12(9):3715–3722. doi: 10.1128/mcb.12.9.3715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fangman W. L., Brewer B. J. Activation of replication origins within yeast chromosomes. Annu Rev Cell Biol. 1991;7:375–402. doi: 10.1146/annurev.cb.07.110191.002111. [DOI] [PubMed] [Google Scholar]
  9. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  10. Ferguson B. M., Brewer B. J., Reynolds A. E., Fangman W. L. A yeast origin of replication is activated late in S phase. Cell. 1991 May 3;65(3):507–515. doi: 10.1016/0092-8674(91)90468-e. [DOI] [PubMed] [Google Scholar]
  11. Forsburg S. L., Nurse P. Cell cycle regulation in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe. Annu Rev Cell Biol. 1991;7:227–256. doi: 10.1146/annurev.cb.07.110191.001303. [DOI] [PubMed] [Google Scholar]
  12. Gahn T. A., Schildkraut C. L. The Epstein-Barr virus origin of plasmid replication, oriP, contains both the initiation and termination sites of DNA replication. Cell. 1989 Aug 11;58(3):527–535. doi: 10.1016/0092-8674(89)90433-9. [DOI] [PubMed] [Google Scholar]
  13. Greenfeder S. A., Newlon C. S. A replication map of a 61-kb circular derivative of Saccharomyces cerevisiae chromosome III. Mol Biol Cell. 1992 Sep;3(9):999–1013. doi: 10.1091/mbc.3.9.999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Harland R. M., Laskey R. A. Regulated replication of DNA microinjected into eggs of Xenopus laevis. Cell. 1980 Oct;21(3):761–771. doi: 10.1016/0092-8674(80)90439-0. [DOI] [PubMed] [Google Scholar]
  15. Heinzel S. S., Krysan P. J., Tran C. T., Calos M. P. Autonomous DNA replication in human cells is affected by the size and the source of the DNA. Mol Cell Biol. 1991 Apr;11(4):2263–2272. doi: 10.1128/mcb.11.4.2263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Heyer W. D., Sipiczki M., Kohli J. Replicating plasmids in Schizosaccharomyces pombe: improvement of symmetric segregation by a new genetic element. Mol Cell Biol. 1986 Jan;6(1):80–89. doi: 10.1128/mcb.6.1.80. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hsiao C. L., Carbon J. High-frequency transformation of yeast by plasmids containing the cloned yeast ARG4 gene. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3829–3833. doi: 10.1073/pnas.76.8.3829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Huberman J. A., Spotila L. D., Nawotka K. A., el-Assouli S. M., Davis L. R. The in vivo replication origin of the yeast 2 microns plasmid. Cell. 1987 Nov 6;51(3):473–481. doi: 10.1016/0092-8674(87)90643-x. [DOI] [PubMed] [Google Scholar]
  19. Huberman J. A., Zhu J. G., Davis L. R., Newlon C. S. Close association of a DNA replication origin and an ARS element on chromosome III of the yeast, Saccharomyces cerevisiae. Nucleic Acids Res. 1988 Jul 25;16(14A):6373–6384. doi: 10.1093/nar/16.14.6373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hyrien O., Méchali M. Plasmid replication in Xenopus eggs and egg extracts: a 2D gel electrophoretic analysis. Nucleic Acids Res. 1992 Apr 11;20(7):1463–1469. doi: 10.1093/nar/20.7.1463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Johnston L. H., Barker D. G. Characterisation of an autonomously replicating sequence from the fission yeast Schizosaccharomyces pombe. Mol Gen Genet. 1987 Apr;207(1):161–164. doi: 10.1007/BF00331504. [DOI] [PubMed] [Google Scholar]
  22. Kearsey S. Structural requirements for the function of a yeast chromosomal replicator. Cell. 1984 May;37(1):299–307. doi: 10.1016/0092-8674(84)90326-x. [DOI] [PubMed] [Google Scholar]
  23. Krysan P. J., Calos M. P. Replication initiates at multiple locations on an autonomously replicating plasmid in human cells. Mol Cell Biol. 1991 Mar;11(3):1464–1472. doi: 10.1128/mcb.11.3.1464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Krysan P. J., Smith J. G., Calos M. P. Autonomous replication in human cells of multimers of specific human and bacterial DNA sequences. Mol Cell Biol. 1993 May;13(5):2688–2696. doi: 10.1128/mcb.13.5.2688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Linskens M. H., Huberman J. A. Ambiguities in results obtained with 2D gel replicon mapping techniques. Nucleic Acids Res. 1990 Feb 11;18(3):647–652. doi: 10.1093/nar/18.3.647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mahbubani H. M., Paull T., Elder J. K., Blow J. J. DNA replication initiates at multiple sites on plasmid DNA in Xenopus egg extracts. Nucleic Acids Res. 1992 Apr 11;20(7):1457–1462. doi: 10.1093/nar/20.7.1457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Marahrens Y., Stillman B. A yeast chromosomal origin of DNA replication defined by multiple functional elements. Science. 1992 Feb 14;255(5046):817–823. doi: 10.1126/science.1536007. [DOI] [PubMed] [Google Scholar]
  28. Martín-Parras L., Hernández P., Martínez-Robles M. L., Schvartzman J. B. Initiation of DNA replication in ColE1 plasmids containing multiple potential origins of replication. J Biol Chem. 1992 Nov 5;267(31):22496–22505. [PubMed] [Google Scholar]
  29. Martín-Parras L., Hernández P., Martínez-Robles M. L., Schvartzman J. B. Unidirectional replication as visualized by two-dimensional agarose gel electrophoresis. J Mol Biol. 1991 Aug 20;220(4):843–853. doi: 10.1016/0022-2836(91)90357-c. [DOI] [PubMed] [Google Scholar]
  30. Maundrell K., Hutchison A., Shall S. Sequence analysis of ARS elements in fission yeast. EMBO J. 1988 Jul;7(7):2203–2209. doi: 10.1002/j.1460-2075.1988.tb03059.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Maundrell K., Wright A. P., Piper M., Shall S. Evaluation of heterologous ARS activity in S. cerevisiae using cloned DNA from S. pombe. Nucleic Acids Res. 1985 May 24;13(10):3711–3722. doi: 10.1093/nar/13.10.3711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Moreno S., Klar A., Nurse P. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol. 1991;194:795–823. doi: 10.1016/0076-6879(91)94059-l. [DOI] [PubMed] [Google Scholar]
  33. Newlon C. S. Yeast chromosome replication and segregation. Microbiol Rev. 1988 Dec;52(4):568–601. doi: 10.1128/mr.52.4.568-601.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Okazaki K., Okazaki N., Kume K., Jinno S., Tanaka K., Okayama H. High-frequency transformation method and library transducing vectors for cloning mammalian cDNAs by trans-complementation of Schizosaccharomyces pombe. Nucleic Acids Res. 1990 Nov 25;18(22):6485–6489. doi: 10.1093/nar/18.22.6485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Palzkill T. G., Oliver S. G., Newlon C. S. DNA sequence analysis of ARS elements from chromosome III of Saccharomyces cerevisiae: identification of a new conserved sequence. Nucleic Acids Res. 1986 Aug 11;14(15):6247–6264. doi: 10.1093/nar/14.15.6247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Prentice H. L. High efficiency transformation of Schizosaccharomyces pombe by electroporation. Nucleic Acids Res. 1992 Feb 11;20(3):621–621. doi: 10.1093/nar/20.3.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Russell P., Nurse P. Schizosaccharomyces pombe and Saccharomyces cerevisiae: a look at yeasts divided. Cell. 1986 Jun 20;45(6):781–782. doi: 10.1016/0092-8674(86)90550-7. [DOI] [PubMed] [Google Scholar]
  38. Sakaguchi J., Yamamoto M. Cloned ural locus of Schizosaccharomyces pombe propagates autonomously in this yeast assuming a polymeric form. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7819–7823. doi: 10.1073/pnas.79.24.7819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sakai K., Sakaguchi J., Yamamoto M. High-frequency cotransformation by copolymerization of plasmids in the fission yeast Schizosaccharomyces pombe. Mol Cell Biol. 1984 Apr;4(4):651–656. doi: 10.1128/mcb.4.4.651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Shinomiya T., Ina S. Analysis of chromosomal replicons in early embryos of Drosophila melanogaster by two-dimensional gel electrophoresis. Nucleic Acids Res. 1991 Jul 25;19(14):3935–3941. doi: 10.1093/nar/19.14.3935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Shinomiya T., Ina S. DNA replication of histone gene repeats in Drosophila melanogaster tissue culture cells: multiple initiation sites and replication pause sites. Mol Cell Biol. 1993 Jul;13(7):4098–4106. doi: 10.1128/mcb.13.7.4098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Stinchcomb D. T., Struhl K., Davis R. W. Isolation and characterisation of a yeast chromosomal replicator. Nature. 1979 Nov 1;282(5734):39–43. doi: 10.1038/282039a0. [DOI] [PubMed] [Google Scholar]
  43. Struhl K., Stinchcomb D. T., Scherer S., Davis R. W. High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1035–1039. doi: 10.1073/pnas.76.3.1035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Symington L. S. Double-strand-break repair and recombination catalyzed by a nuclear extract of Saccharomyces cerevisiae. EMBO J. 1991 Apr;10(4):987–996. doi: 10.1002/j.1460-2075.1991.tb08033.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tran C. T., Caddle M. S., Calos M. P. The replication behavior of Saccharomyces cerevisiae DNA in human cells. Chromosoma. 1993 Jan;102(2):129–136. doi: 10.1007/BF00356030. [DOI] [PubMed] [Google Scholar]
  46. Umek R. M., Linskens M. H., Kowalski D., Huberman J. A. New beginnings in studies of eukaryotic DNA replication origins. Biochim Biophys Acta. 1989 Jan 23;1007(1):1–14. doi: 10.1016/0167-4781(89)90123-1. [DOI] [PubMed] [Google Scholar]
  47. Van Houten J. V., Newlon C. S. Mutational analysis of the consensus sequence of a replication origin from yeast chromosome III. Mol Cell Biol. 1990 Aug;10(8):3917–3925. doi: 10.1128/mcb.10.8.3917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Vaughn J. P., Dijkwel P. A., Hamlin J. L. Replication initiates in a broad zone in the amplified CHO dihydrofolate reductase domain. Cell. 1990 Jun 15;61(6):1075–1087. doi: 10.1016/0092-8674(90)90071-l. [DOI] [PubMed] [Google Scholar]
  49. Wellinger R. J., Wolf A. J., Zakian V. A. Origin activation and formation of single-strand TG1-3 tails occur sequentially in late S phase on a yeast linear plasmid. Mol Cell Biol. 1993 Jul;13(7):4057–4065. doi: 10.1128/mcb.13.7.4057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. 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]
  51. Yang L., Botchan M. Replication of bovine papillomavirus type 1 DNA initiates within an E2-responsive enhancer element. J Virol. 1990 Dec;64(12):5903–5911. doi: 10.1128/jvi.64.12.5903-5911.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Zhu J., Brun C., Kurooka H., Yanagida M., Huberman J. A. Identification and characterization of a complex chromosomal replication origin in Schizosaccharomyces pombe. Chromosoma. 1992;102(1 Suppl):S7–16. doi: 10.1007/BF02451780. [DOI] [PubMed] [Google Scholar]
  53. Zhu J., Newlon C. S., Huberman J. A. Localization of a DNA replication origin and termination zone on chromosome III of Saccharomyces cerevisiae. Mol Cell Biol. 1992 Oct;12(10):4733–4741. doi: 10.1128/mcb.12.10.4733. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES