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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 Dec;79(24):7819–7823. doi: 10.1073/pnas.79.24.7819

Cloned ural locus of Schizosaccharomyces pombe propagates autonomously in this yeast assuming a polymeric form.

J Sakaguchi, M Yamamoto
PMCID: PMC347440  PMID: 6961452

Abstract

DNA segments cloned from Schizosaccharomyces pombe by the ability to complement Escherichia coli pyrB mutations are shown to complement a ural mutation in S. pombe, thereby demonstrating that ural is the structural gene for aspartate transcarbamylase of S. pombe. Further, such segments combined with parts or all of pBR322 are shown to be capable of autonomous propagation in S. pombe. This suggests the existence of an autonomously replicating sequence (ars) in the vicinity of ural. Unlike the TRP1 segment cloned from Saccharomyces cerevisiae [Struhl, K., Stinchcomb, D. T., Scherer, S. & Davis, R. W. (1979) Proc. Natl. Acad. Sci. USA 76, 1035-1039], plasmids carrying the ural locus do not multiply as monomers but assume a polymeric form as large as a decamer to an icosamer in the yeast. Monomers are tandemly arranged in the polymer. Inversion of an inserted fragment or insertion of another segment into a competent plasmid greatly decreases the efficiency of such transformation, implying a role of the tertiary structure of the plasmids in the establishment of transformation of this kind.

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Selected References

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  1. Beggs J. D. Transformation of yeast by a replicating hybrid plasmid. Nature. 1978 Sep 14;275(5676):104–109. doi: 10.1038/275104a0. [DOI] [PubMed] [Google Scholar]
  2. Blanc H., Dujon B. Replicator regions of the yeast mitochondrial DNA responsible for suppressiveness. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3942–3946. doi: 10.1073/pnas.77.7.3942. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Borst P., Hoeijmakers J. H. Kinetoplast DNA. Plasmid. 1979 Jan;2(1):20–40. doi: 10.1016/0147-619x(79)90003-9. [DOI] [PubMed] [Google Scholar]
  4. Botstein D., Falco S. C., Stewart S. E., Brennan M., Scherer S., Stinchcomb D. T., Struhl K., Davis R. W. Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments. Gene. 1979 Dec;8(1):17–24. doi: 10.1016/0378-1119(79)90004-0. [DOI] [PubMed] [Google Scholar]
  5. Clarke L., Carbon J. Isolation of a yeast centromere and construction of functional small circular chromosomes. Nature. 1980 Oct 9;287(5782):504–509. doi: 10.1038/287504a0. [DOI] [PubMed] [Google Scholar]
  6. Clayton D. A., Vinograd J. Circular dimer and catenate forms of mitochondrial DNA in human leukaemic leucocytes. Nature. 1967 Nov 18;216(5116):652–657. doi: 10.1038/216652a0. [DOI] [PubMed] [Google Scholar]
  7. Cryer D. R., Eccleshall R., Marmur J. Isolation of yeast DNA. Methods Cell Biol. 1975;12:39–44. doi: 10.1016/s0091-679x(08)60950-4. [DOI] [PubMed] [Google Scholar]
  8. Daniels D. L., de Wet J. R., Blattner F. R. New map of bacteriophage lambda DNA. J Virol. 1980 Jan;33(1):390–400. doi: 10.1128/jvi.33.1.390-400.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Del Giudice L., Wolf K., Sassone-Corsi P., Mazza A. 2 micrometer covalently closed non-mitochondrial circular DNA in the petite-negative yeast Schizosaccharomyces pombe. Mol Gen Genet. 1979 May 4;172(2):165–169. doi: 10.1007/BF00268278. [DOI] [PubMed] [Google Scholar]
  10. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  11. Egel R., Kohli J., Thuriaux P., Wolf K. Genetics of the fission yeast Schizosaccharomyces pombe. Annu Rev Genet. 1980;14:77–108. doi: 10.1146/annurev.ge.14.120180.000453. [DOI] [PubMed] [Google Scholar]
  12. Hinnen A., Hicks J. B., Fink G. R. Transformation of yeast. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929–1933. doi: 10.1073/pnas.75.4.1929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Holland M. J., Holland J. P., Jackson K. A. Cloning of yeast genes coding for glycolytic enzymes. Methods Enzymol. 1979;68:408–419. doi: 10.1016/0076-6879(79)68030-8. [DOI] [PubMed] [Google Scholar]
  14. Hudson B., Vinograd J. Catenated circular DNA molecules in HeLa cell mitochondria. Nature. 1967 Nov 18;216(5116):647–652. doi: 10.1038/216647a0. [DOI] [PubMed] [Google Scholar]
  15. Kohli J., Hottinger H., Munz P., Strauss A., Thuriaux P. Genetic Mapping in SCHIZOSACCHAROMYCES POMBE by Mitotic and Meiotic Analysis and Induced Haploidization. Genetics. 1977 Nov;87(3):471–489. doi: 10.1093/genetics/87.3.471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mitchison J. M., Creanor J. Further measurements of DNA synthesis and enzyme potential during cell cycle of fission yeast Schizosaccharomyces pombe. Exp Cell Res. 1971 Nov;69(1):244–247. doi: 10.1016/0014-4827(71)90337-5. [DOI] [PubMed] [Google Scholar]
  17. Perkins D. D. Biochemical Mutants in the Smut Fungus Ustilago Maydis. Genetics. 1949 Sep;34(5):607–626. doi: 10.1093/genetics/34.5.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  19. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  20. Stinchcomb D. T., Mann C., Davis R. W. Centromeric DNA from Saccharomyces cerevisiae. J Mol Biol. 1982 Jun 25;158(2):157–190. doi: 10.1016/0022-2836(82)90427-2. [DOI] [PubMed] [Google Scholar]
  21. Struhl K., Davis R. W. Position effects in Saccharomyces cerevisiae. J Mol Biol. 1981 Nov 5;152(3):569–575. doi: 10.1016/0022-2836(81)90269-2. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Stüber D., Bujard H. Organization of transcriptional signals in plasmids pBR322 and pACYC184. Proc Natl Acad Sci U S A. 1981 Jan;78(1):167–171. doi: 10.1073/pnas.78.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Yang R., Lis J., Wu R. Elution of DNA from agarose gels after electrophoresis. Methods Enzymol. 1979;68:176–182. doi: 10.1016/0076-6879(79)68012-6. [DOI] [PubMed] [Google Scholar]

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