Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1984 Sep;4(9):1725–1729. doi: 10.1128/mcb.4.9.1725

Micronuclear DNA of Oxytricha nova contains sequences with autonomously replicating activity in Saccharomyces cerevisiae.

M M Colombo, M T Swanton, P Donini, D M Prescott
PMCID: PMC368979  PMID: 6092934

Abstract

Oxytricha nova is a hypotrichous ciliate with micronuclei and macronuclei. Micronuclei, which contain large, chromosomal-sized DNA, are genetically inert but undergo meiosis and exchange during cell mating. Macronuclei, which contain only small, gene-sized DNA molecules, provide all of the nuclear RNA needed to run the cell. After cell mating the macronucleus is derived from a micronucleus, a derivation that includes excision of the genes from chromosomes and elimination of the remaining DNA. The eliminated DNA includes all of the repetitious sequences and approximately 95% of the unique sequences. We cloned large restriction fragments from the micronucleus that confer replication ability on a replication-deficient plasmid in Saccharomyces cerevisiae. Sequences that confer replication ability are called autonomously replicating sequences. The frequency and effectiveness of autonomously replicating sequences in micronuclear DNA are similar to those reported for DNAs of other organisms introduced into yeast cells. Of the 12 micronuclear fragments with autonomously replicating sequence activity, 9 also showed homology to macronuclear DNA, indicating that they contain a macronuclear gene sequence. We conclude from this that autonomously replicating sequence activity is nonrandomly distributed throughout micronuclear DNA and is preferentially associated with those regions of micronuclear DNA that contain genes.

Full text

PDF
1725

Images in this article

Selected References

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

  1. Boswell R. E., Jahn C. L., Greslin A. F., Prescott D. M. Organization of gene and non-gene sequences in micronuclear DNA of Oxytricha nova. Nucleic Acids Res. 1983 Jun 11;11(11):3651–3663. doi: 10.1093/nar/11.11.3651. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boswell R. E., Klobutcher L. A., Prescott D. M. Inverted terminal repeats are added to genes during macronuclear development in Oxytricha nova. Proc Natl Acad Sci U S A. 1982 May;79(10):3255–3259. doi: 10.1073/pnas.79.10.3255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bouché J. P. The effect of spermidine on endonuclease inhibition by agarose contaminants. Anal Biochem. 1981 Jul 15;115(1):42–45. doi: 10.1016/0003-2697(81)90519-4. [DOI] [PubMed] [Google Scholar]
  4. Burke W., Fangman W. L. Temporal order in yeast chromosome replication. Cell. 1975 Jul;5(3):263–269. doi: 10.1016/0092-8674(75)90101-4. [DOI] [PubMed] [Google Scholar]
  5. Cameron J. R., Philippsen P., Davis R. W. Analysis of chromosomal integration and deletions of yeast plasmids. Nucleic Acids Res. 1977;4(5):1429–1448. doi: 10.1093/nar/4.5.1429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chan C. S., Tye B. K. Autonomously replicating sequences in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6329–6333. doi: 10.1073/pnas.77.11.6329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clewell D. B. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol. J Bacteriol. 1972 May;110(2):667–676. doi: 10.1128/jb.110.2.667-676.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Goursot R., Goze A., Niaudet B., Ehrlich S. D. Plasmids from Staphylococcus aureus replicate in yeast Saccharomyces cerevisiae. Nature. 1982 Jul 29;298(5873):488–490. doi: 10.1038/298488a0. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  13. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Loening U. E. The determination of the molecular weight of ribonucleic acid by polyacrylamide-gel electrophresis. The effects of changes in conformation. Biochem J. 1969 Jun;113(1):131–138. doi: 10.1042/bj1130131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Murti K. G., Prescott D. M. Replication forms of the gene-sized DNA molecules of hypotrichous ciliates. Mol Cell Biol. 1983 Sep;3(9):1562–1566. doi: 10.1128/mcb.3.9.1562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Roth G. E., Blanton H. M., Hager L. J., Zakian V. A. Isolation and characterization of sequences from mouse chromosomal DNA with ARS function in yeasts. Mol Cell Biol. 1983 Nov;3(11):1898–1908. doi: 10.1128/mcb.3.11.1898. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Spear B. B., Lauth M. R. Polytene chromosomes of Oxytricha: biochemical and morphological changes during macronuclear development in a ciliated protozoan. Chromosoma. 1976 Jan 27;54(1):1–13. doi: 10.1007/BF00331828. [DOI] [PubMed] [Google Scholar]
  20. Stinchcomb D. T., Thomas M., Kelly J., Selker E., Davis R. W. Eukaryotic DNA segments capable of autonomous replication in yeast. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4559–4563. doi: 10.1073/pnas.77.8.4559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Swanton M. T., Greslin A. F., Prescott D. M. Arrangement of coding and non-coding sequences in the DNA molecules coding for rRNAs in Oxytricha sp. DNA of ciliated protozoa. VII. Chromosoma. 1980;77(2):203–215. doi: 10.1007/BF00329545. [DOI] [PubMed] [Google Scholar]
  23. Tschumper G., Carbon J. Sequence of a yeast DNA fragment containing a chromosomal replicator and the TRP1 gene. Gene. 1980 Jul;10(2):157–166. doi: 10.1016/0378-1119(80)90133-x. [DOI] [PubMed] [Google Scholar]
  24. Zyskind J. W., Cleary J. M., Brusilow W. S., Harding N. E., Smith D. W. Chromosomal replication origin from the marine bacterium Vibrio harveyi functions in Escherichia coli: oriC consensus sequence. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1164–1168. doi: 10.1073/pnas.80.5.1164. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES