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Survival Strategies of the Yeast Plasmid Two-Micron Circle

  • Fredric C. Volkert
  • Ling-Chuan Chen Wu
  • Paul A. Fisher
  • James R. Broach
Part of the Basic Life Sciences book series (BLSC, volume 40)

Abstract

The multicopy yeast plasmid 2-micron circle uses a number of strategies to insure its persistence in its host. The plasmid confers no selective phenotype to the cell in which it is resident. Nonetheless, the plasmid is lost at less than 1 per 105 cell divisions during continuous exponential growth. We have determined that the plasmid persists at least in part due to the ability of the plasmid to amplify its mean copy number when its cellular copy level is low and to distribute plasmid molecules equally between mother and daughter cells at mitosis. We have found that amplification of plasmid copy number occurs by a novel mechanism in which site-specific recombination induces a transient shift in the mode of replication from theta to rolling circle. Equitable partitioning of plasmid molecules requires plasmid-encoded proteins and a centromere-like segment on the plasmid. We have accumulated evidence consistent with a model of partitioning in which the partitioning proteins form a transnuclear structure that is responsible for distributing plasmid molecules throughout the nucleus prior to cell division. In this chapter we describe evidence supporting the existence and mode of action of these two plasmid strategies and discuss the extent to which these strategies may be a pervasive facet of the biology of eukaryotic extrachromosomal elements.

Keywords

Nuclear Matrix REP1 Protein Nuclear Pore Complex Plasmid Copy Plasmid Copy Number 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Aaronson, R.P., and G. Blobel (1975) Isolation of nuclear pore complexes in association with a lamina. Proc. Natl. Acad. Sci., USA 72:1007–1011.PubMedCrossRefGoogle Scholar
  2. 2.
    Adams, B.G. (1972) Induction of galactokinase in Saccharomyces cerevisiae; Kinetics of induction and glucose effects. J. Bacteriol. 111:308–315.PubMedGoogle Scholar
  3. 3.
    Agutter, P.S., and J.C.W. Richardson (1980) Nuclear non-chromatin proteinaceous structures: Their role in the organization and function of the interphase nucleus. J. Cell Sci. 44:395–435.PubMedGoogle Scholar
  4. 4.
    Andrews, B.J., G.A. Proteau, L.G. Beatty, and P.D. Sadowski (1985) The FLP recombinase of the 2 micron circle DNA of yeast: Interaction with its target sequences. Cell 40:795–803.PubMedCrossRefGoogle Scholar
  5. 5.
    Araki, H., A. Jearnpipatkul, H. Tatsumi, T. Sakurai, K. Ushio, T. Muta, and Y. Oshima (1985) Molecular and functional organization of yeast plasmid pSR1. J. Mol. Biol. 182:191–203.PubMedCrossRefGoogle Scholar
  6. 6.
    Berezney, R. (1984) Organization and functions of the nuclear matrix. In Chromosomal Nonhistone Proteins, Vol. 4, L.S. Hnilica, ed. CRC Press, Inc., Boca Raton, Florida, pp. 119–180.Google Scholar
  7. 7.
    Berezney, R., and D.S. Coffey (1977) Nuclear matrix. Isolation and characterization of a framework structure from rat liver nuclei. J. Cell Biol. 73:616–637.PubMedCrossRefGoogle Scholar
  8. 8.
    Berrios, M., and P.A. Fisher (1986) A myosin heavy chain-like polypeptide is associated with the nuclear envelope in higher eukaryotic cells. J. Cell Biol. (in press).Google Scholar
  9. 9.
    Berrios, M., G. Blobel, and P.A. Fisher (1983) Characterization of an ATPase/dATPase activity associated with the Drosophila nuclear matrix-pore complex-lamina fraction. Identification of the putative enzyme polypeptide by direct ultraviolet photoaffinity labeling. J. Biol. Chem. 258:4548–4555.PubMedGoogle Scholar
  10. 10.
    Bouteille, M., D. Bouvier, and A.P. Seve (1983) Heterogeneity and territorial organization of the nuclear matrix and related structures. Int. Rev. Cytol. 83:135–182.PubMedCrossRefGoogle Scholar
  11. 11.
    Broach, J.R. (1981) The yeast plasmid 2 micron circle. In The Molecular Biology of the Yeast Saccharomyces: Life Cycle and Inheritance, J.N. Strathern, E.W. Jones, and J.R. Broach, eds. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp. 445–470.Google Scholar
  12. 12.
    Broach, J.R., and J.B. Hicks (1980) Replication and recombination functions associated with the yeast plasmid 2 micron circle. Cell 21:501–508.PubMedCrossRefGoogle Scholar
  13. 13.
    Broach, J.R., V.R. Guarascio, and M. Jayaram (1982) Recombination within the yeast plasmid 2 micron circle is site-specific. Cell 29: 227–234.PubMedCrossRefGoogle Scholar
  14. 14.
    Broach, J.R., Y.-Y. Li, L.-C.C. Wu, and M. Jayaram (1983) Vectors for high-level, inducible expression of cloned genes in yeast. In Experimental Manipulation of Gene Expression, M. Inouye, ed. Academic Press, Inc., New York, pp. 83–117.Google Scholar
  15. 15.
    Burke, B., and L. Gerace (1986) A cell free system to study reassembly of the nuclear envelope at the end of mitosis. Cell 44:639–652.PubMedCrossRefGoogle Scholar
  16. 16.
    Conde, J., and G.R. Fink (1976) A mutant of Saccharomyces cerevisiae defective for nuclear fusion. Proc. Natl. Acad. Sci., USA 73:3651–3655.PubMedCrossRefGoogle Scholar
  17. 17.
    Cox, M.M. (1983) The FLP protein of the yeast 2-micron plasmid: Expression of a eukaryotic genetic recombination system in Escherichia coli. Proc. Natl. Acad. Sci., USA 80:4223–4227.PubMedCrossRefGoogle Scholar
  18. 18.
    Dwyer, N., and G. Blobel (1976) A modified procedure for isolation of a pore complex-lamina fraction from rat liver nuclei. J. Cell Biol. 70:581–591.PubMedCrossRefGoogle Scholar
  19. 19.
    Filson, A.J., A. Lewis, G. Blobel, and P.A. Fisher (1985) Monoclonal antibodies prepared against the major Drosophila nuclear matrix-pore complex-lamina glycoprotein bind specifically to the nuclear envelope in situ. J. Biol. Chem. 260:3164–3172.PubMedGoogle Scholar
  20. 20.
    Fisher, P.A. (1986) Karyoskeletal proteins of Drosophila. In Chromosome and Chromatin Structure, K.W. Adolph, ed. CRC Press, Inc., Boca Raton, Florida (in press).Google Scholar
  21. 21.
    Fisher, P.A., M. Berrios, and G. Blobel (1982) Isolation and characterization of a proteinaceous subnuclear fraction composed of nuclear matrix, peripheral lamina, and nuclear pore complexes from embryos of Drosophila melanogaster. J. Cell Biol. 92:674–686.PubMedCrossRefGoogle Scholar
  22. 22.
    Futcher, A.B. (1986) Copy number amplification of the 2 micron circle plasmid of Saccharomyces cerevisiae. J. Theoret. Biol. (in press).Google Scholar
  23. 23.
    Futcher, A.B., and B.S. Cox (1983) Maintenance of the 2 micron circle plasmid in populations of Saccharomyces cerevisiae. J. Bacteriol. 154:612–622.PubMedGoogle Scholar
  24. 24.
    Gerace, L., Y. Ottaviano, and C. Kondor-Koch (1982) Identification of a major polypeptide of the nuclear pore complex. J. Cell Biol. 95: 826–837.PubMedCrossRefGoogle Scholar
  25. 25.
    Gunge, N. (1983) Yeast DNA plasmids. Ann. Rev. Microbiol. 37:253.CrossRefGoogle Scholar
  26. 26.
    Hartley, J.L., and J.E. Donelson (1980) Nucleotide sequence of the yeast plasmid. Nature 286:860–865.PubMedCrossRefGoogle Scholar
  27. 27.
    Jayaram, M., Y.-Y. Li, and J.R. Broach (1983) The yeast plasmid 2 micron circle encodes components required for its high copy propagation. Cell 34:95–104.PubMedCrossRefGoogle Scholar
  28. 28.
    Jayaram, M., A. Sutton, and J.R. Broach (1985) Properties of REP3: A cis acting locus required for stable propagation of the yeast plasmid 2 micron circle. Mol. Cell. Biol. 5:2466.PubMedGoogle Scholar
  29. 29.
    Kay, R.R., D. Fraser, and I.R. Johnston (1972) A method for the rapid isolation of nuclear membranes from rat liver. Characterization of the membrane preparation and its associated DNA polymerase. Eur. J. Biochem. 30:145–154.PubMedCrossRefGoogle Scholar
  30. 30.
    Kikuchi, Y. (1983) Yeast plasmid requires a cis-acting locus and two plasmid proteins for its stable maintenance. Cell 35:487–493.PubMedCrossRefGoogle Scholar
  31. 31.
    Krohne, G., W.W. Franke, and U. Scheer (1978) The major polypeptides of the nuclear pore complex. Exp. Cell Res. 116:85–102.PubMedCrossRefGoogle Scholar
  32. 32.
    Lipman, D.J., and W.R. Pearson (1985) Rapid and sensitive protein similarity searches. Science 227:1435–1441.PubMedCrossRefGoogle Scholar
  33. 33.
    Livingston, D.M. (1977) Inheritance of 2 micron DNA from Saccharomyces. Genetics 86:73–82.PubMedGoogle Scholar
  34. 34.
    Livingston, D.M., and D.M. Kupfer (1977) Control of Saccharomyces cerevisiae 2 micron DNA replication by cell division cycle genes that control nuclear DNA replication. J. Mol. Biol. 116:249–260.PubMedCrossRefGoogle Scholar
  35. 35.
    Maine, G.T., P. Sinha, and B.-K. Tye (1984) Mutants of S. cerevisiae defective in the maintenance of minichromosomes. Genetics 106:365–375.PubMedGoogle Scholar
  36. 36.
    McLeod, M., F. Volkert, and J. Broach (1984) Components of the site-specific recombination system encoded by the yeast plasmid 2-micron circle. Cold Spring Harbor Symp. Quant. Biol. 49:779–787.PubMedCrossRefGoogle Scholar
  37. 37.
    McLeod, M., S. Craft, and J.R. Broach (1986) Identification of the crossover site during FLP-mediated recombination in the yeast plasmid 2 micron circle. Mol. Cell. Biol. (in press).Google Scholar
  38. 38.
    Mortimer, R.K., and J.R. Johnston (1959) Life span of individual yeast cells. Nature 183:1751–1752.PubMedCrossRefGoogle Scholar
  39. 39.
    Murray, A.W., and J.W. Szostak (1983) Pedigree analysis of plasmid segregation in yeast. Cell 34:961–970.PubMedCrossRefGoogle Scholar
  40. 40.
    Newlon, C.S., R.J. Devenish, R.A. Suci, and C.J. Roffis (1981) Replication origins used in vivo in yeast. In Structure and DNA Protein Interactions of Replication Origins (ICN-UCLA Symposia), D.S. Ray and C.F. Fox, eds. Mol. Cell. Biol. 22:501–516.Google Scholar
  41. 41.
    Pienta, K., and D. Coffey (1984) A structural analysis of the role of the nuclear matrix and DNA loops in the organization of the nucleus and chromosome. J. Cell Sci. Supplement 1, pp. 123–135.Google Scholar
  42. 42.
    Rohlf, F.J., and R.R. Sokal (1969) Statistical Tables, W.H. Freeman, San Francisco, p. 214.Google Scholar
  43. 43.
    Sigurdson, D.C., M.E. Gaarder, and D.M. Livingston (1981) Characterization of the transmission during cytoductant formation of the 2 micron DNA plasmid from Saccharomyces. Mol. Gen. Genet. 183:59–65.PubMedCrossRefGoogle Scholar
  44. 44.
    Southern, E.M. (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98:503–517.PubMedCrossRefGoogle Scholar
  45. 45.
    Steinert, P.M., A.C. Stevens, and D.R. Roop (1985) The molecular biology of intermediate filaments. Cell 42:411–419.PubMedCrossRefGoogle Scholar
  46. 46.
    Tohe, A., S. Tada, and Y. Oshima (1982) Two-micron DNA-like plasmids in the osmophilic haploid yeast Saccharomyces rouxii. J. Bacteriol. 151:1380–1390.Google Scholar
  47. 47.
    Tohe, A., H. Araki, I. Utatsu, and Y. Oshima (1984) Plasmids resembling 2-micron DNA in the osmotolerant yeasts Saccharomyces bailii and Saccharomyces bisporus. J. Gen. Microbiol. 130:2527–2534.Google Scholar
  48. 48.
    Veit, B.E., and W.L. Fangman (1985) Chromatin organization of the Saccharomyces cerevisiae 2 micron plasmid depends on plasmid-encoded products. Mol. Cell. Biol. 5:2190–2196.PubMedGoogle Scholar
  49. 49.
    Vetter, D., B.J. Andrews, L. Roberts-Beatty, and P.D. Sadowski (1983) Site-specific recombination of yeast 2-micron DNA in vitro. Proc. Natl. Acad. Sci., USA 80:7284–7288.PubMedCrossRefGoogle Scholar
  50. 50.
    Volkert, F.C., and J.R. Broach (1987) The mechanism of propagation of the yeast 2 micron circle plasmid. In Industrial Biochemistry and Molecular Biology of Yeasts, G.G. Stewart, I. Russell, R.D. Klein, and R.R. Hiebisch, eds. CRC Press, Inc., Boca Raton, Florida (in press).Google Scholar
  51. 51.
    Volkert, F.C., and J.R. Broach (1986) Site-specific recombination promotes plasmid amplification in yeast. Cell (in press).Google Scholar
  52. 52.
    Williamson, D.H. (1985) The yeast ARS element, six years on: A progress report. Yeast 1:1–14.PubMedCrossRefGoogle Scholar
  53. 53.
    Wu, L.C.C., P.A. Fisher, and J.R. Broach (1986) The REP1 protein of 2 micron circle is associated with the yeast nuclear matrix. In Yeast Cell Biology, J.B. Hicks and C.F. Fox, eds. Academic Press, Inc., New York (in press).Google Scholar
  54. 54.
    Wu, L.C.C., P.A. Fisher, and J.R. Broach (1986) The REP1 protein of yeast is a karyoskeletal component. J. Biol. Chem. (submitted for publication).Google Scholar
  55. 55.
    Zakian, V.A., B.J. Brewer, and W.L. Fangman (1979) Replication of each copy of the yeast 2 micron DNA plasmid occurs during the S phase. Cell 17:923–934.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1986

Authors and Affiliations

  • Fredric C. Volkert
    • 1
  • Ling-Chuan Chen Wu
    • 2
  • Paul A. Fisher
    • 2
  • James R. Broach
    • 1
  1. 1.Department of Molecular Biology, Lewis Thomas LaboratoryPrinceton UniversityPrincetonUSA
  2. 2.Department of Pharmacology, Health Sciences CenterState University of New YorkStony BrookUSA

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