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. 1992 Sep;3(9):999–1013. doi: 10.1091/mbc.3.9.999

A replication map of a 61-kb circular derivative of Saccharomyces cerevisiae chromosome III.

S A Greenfeder 1, C S Newlon 1
PMCID: PMC275661  PMID: 1330093

Abstract

Using two-dimensional agarose gel electrophoresis, we determined the replication map of a 61-kb circular derivative of Saccharomyces cerevisiae chromosome III. The three sites of DNA replication initiation on the ring chromosome are specific and coincide with ARS elements. The three origins are active to different degrees; two are used > 90% of the time, whereas the third is used only 10-20% of the time. The specificity of these origins is shown by the fact that only ARS elements were competent for origin function, and deletion of one of the ARS elements removed the corresponding replication origin. The activity of the least active origin was not increased by deletion of the nearby highly active origin, demonstrating that the highly active origin does not repress function of the relatively inactive origin. Replication termination on the ring chromosome does not occur at specific sites but rather occurs over stretches of DNA ranging from 3 to 10 kb. A new region of termination was created by altering the sites of initiation. The position of the new termination site indicates that termination is not controlled by specific cis-acting DNA sequences, but rather that replication termination is determined primarily by the positions at which replication initiates. In addition, two sites on the ring chromosome were found to slow the progression of replication forks through the molecule: one is at the centromere and one at the 3' end of a yeast transposable element.

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Selected References

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

  1. Boeke J. D., Trueheart J., Natsoulis G., Fink G. R. 5-Fluoroorotic acid as a selective agent in yeast molecular genetics. Methods Enzymol. 1987;154:164–175. doi: 10.1016/0076-6879(87)54076-9. [DOI] [PubMed] [Google Scholar]
  2. Brewer B. J., Fangman W. L. A replication fork barrier at the 3' end of yeast ribosomal RNA genes. Cell. 1988 Nov 18;55(4):637–643. doi: 10.1016/0092-8674(88)90222-x. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Brewer B. J. When polymerases collide: replication and the transcriptional organization of the E. coli chromosome. Cell. 1988 Jun 3;53(5):679–686. doi: 10.1016/0092-8674(88)90086-4. [DOI] [PubMed] [Google Scholar]
  5. Brockman W. W., Gutai M. W., Nathans D. Evolutionary variants of simian virus 40: characterization of cloned complementing variants. Virology. 1975 Jul;66(1):36–52. doi: 10.1016/0042-6822(75)90177-4. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Dubey D. D., Davis L. R., Greenfeder S. A., Ong L. Y., Zhu J. G., Broach J. R., Newlon C. S., Huberman J. A. Evidence suggesting that the ARS elements associated with silencers of the yeast mating-type locus HML do not function as chromosomal DNA replication origins. Mol Cell Biol. 1991 Oct;11(10):5346–5355. doi: 10.1128/mcb.11.10.5346. [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. 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]
  10. Ferguson B. M., Fangman W. L. A position effect on the time of replication origin activation in yeast. Cell. 1992 Jan 24;68(2):333–339. doi: 10.1016/0092-8674(92)90474-q. [DOI] [PubMed] [Google Scholar]
  11. Fitzgerald-Hayes M., Clarke L., Carbon J. Nucleotide sequence comparisons and functional analysis of yeast centromere DNAs. Cell. 1982 May;29(1):235–244. doi: 10.1016/0092-8674(82)90108-8. [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. Replication forks pause at yeast centromeres. Mol Cell Biol. 1992 Sep;12(9):4056–4066. doi: 10.1128/mcb.12.9.4056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Griffin B. E., Fried M. Amplification of a specific region of the polyoma virus genome. Nature. 1975 Jul 17;256(5514):175–179. doi: 10.1038/256175a0. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. 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]
  18. Johnston L. H., Williamson D. H. An alkaline sucrose gradient analysis of the mechanism of nuclear DNA synthesis in the yeast Saccharomyces cerevisiae. Mol Gen Genet. 1978 Aug 17;164(2):217–225. doi: 10.1007/BF00267387. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Kuempel P. L., Pelletier A. J., Hill T. M. Tus and the terminators: the arrest of replication in prokaryotes. Cell. 1989 Nov 17;59(4):581–583. doi: 10.1016/0092-8674(89)90001-9. [DOI] [PubMed] [Google Scholar]
  21. Levitt M. Real-time interactive frequency filtering of molecular dynamics trajectories. J Mol Biol. 1991 Jul 5;220(1):1–4. doi: 10.1016/0022-2836(91)90373-e. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Linskens M. H., Huberman J. A. Organization of replication of ribosomal DNA in Saccharomyces cerevisiae. Mol Cell Biol. 1988 Nov;8(11):4927–4935. doi: 10.1128/mcb.8.11.4927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Martin M. A., Khoury G., Fareed G. C. Specific reiteration of viral DNA sequences in mammalian cells. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):129–136. doi: 10.1101/sqb.1974.039.01.018. [DOI] [PubMed] [Google Scholar]
  25. McCarroll R. M., Fangman W. L. Time of replication of yeast centromeres and telomeres. Cell. 1988 Aug 12;54(4):505–513. doi: 10.1016/0092-8674(88)90072-4. [DOI] [PubMed] [Google Scholar]
  26. Mirkovitch J., Mirault M. E., Laemmli U. K. Organization of the higher-order chromatin loop: specific DNA attachment sites on nuclear scaffold. Cell. 1984 Nov;39(1):223–232. doi: 10.1016/0092-8674(84)90208-3. [DOI] [PubMed] [Google Scholar]
  27. Nawotka K. A., Huberman J. A. Two-dimensional gel electrophoretic method for mapping DNA replicons. Mol Cell Biol. 1988 Apr;8(4):1408–1413. doi: 10.1128/mcb.8.4.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Newlon C. S., Lipchitz L. R., Collins I., Deshpande A., Devenish R. J., Green R. P., Klein H. L., Palzkill T. G., Ren R. B., Synn S. Analysis of a circular derivative of Saccharomyces cerevisiae chromosome III: a physical map and identification and location of ARS elements. Genetics. 1991 Oct;129(2):343–357. doi: 10.1093/genetics/129.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Oliver S. G., van der Aart Q. J., Agostoni-Carbone M. L., Aigle M., Alberghina L., Alexandraki D., Antoine G., Anwar R., Ballesta J. P., Benit P. The complete DNA sequence of yeast chromosome III. Nature. 1992 May 7;357(6373):38–46. doi: 10.1038/357038a0. [DOI] [PubMed] [Google Scholar]
  31. Palzkill T. G., Newlon C. S. A yeast replication origin consists of multiple copies of a small conserved sequence. Cell. 1988 May 6;53(3):441–450. doi: 10.1016/0092-8674(88)90164-x. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Rivier D. H., Rine J. An origin of DNA replication and a transcription silencer require a common element. Science. 1992 May 1;256(5057):659–663. doi: 10.1126/science.1585179. [DOI] [PubMed] [Google Scholar]
  34. Saffer L. D., Miller O. L., Jr Electron microscopic study of Saccharomyces cerevisiae rDNA chromatin replication. Mol Cell Biol. 1986 Apr;6(4):1148–1157. doi: 10.1128/mcb.6.4.1148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Strathern J. N., Newlon C. S., Herskowitz I., Hicks J. B. Isolation of a circular derivative of yeast chromosome III: implications for the mechanism of mating type interconversion. Cell. 1979 Oct;18(2):309–319. doi: 10.1016/0092-8674(79)90050-3. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Surosky R. T., Tye B. K. Resolution of dicentric chromosomes by Ty-mediated recombination in yeast. Genetics. 1985 Jul;110(3):397–419. doi: 10.1093/genetics/110.3.397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Szostak J. W., Wu R. Insertion of a genetic marker into the ribosomal DNA of yeast. Plasmid. 1979 Oct;2(4):536–554. doi: 10.1016/0147-619x(79)90053-2. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. 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]
  41. Walker S. S., Malik A. K., Eisenberg S. Analysis of the interactions of functional domains of a nuclear origin of replication from Saccharomyces cerevisiae. Nucleic Acids Res. 1991 Nov 25;19(22):6255–6262. doi: 10.1093/nar/19.22.6255. [DOI] [PMC free article] [PubMed] [Google Scholar]

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