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. 1987 Sep 11;15(17):6937–6956. doi: 10.1093/nar/15.17.6937

Fine analysis of the chromatin structure of the yeast SUC2 gene and of its changes upon derepression. Comparison between the chromosomal and plasmid-inserted genes.

J E Pérez-Ortín 1, F Estruch 1, E Matallana 1, L Franco 1
PMCID: PMC306185  PMID: 2821486

Abstract

Micrococcal nuclease digestion has been used to investigate some fine details of the chromatin structure of the yeast SUC2 gene for invertase. Precisely positioned nucleosomes have been found on a 2 kb sequence from the 3' non-coding region, and four nucleosomes also seem to occupy fixed positions on the 5' flank. Eleven nucleosomes lie on the coding region, although their positioning is not as precise as in the flanks. When the gene is derepressed, these latter nucleosomes adopt a more open conformation and so do two of the nucleosomes positioned on the 5' flank. A dramatic change occurs in the 3' flank, whose involvement in the structural transitions of chromatin upon gene activation is postulated. All the observed features are conserved when the gene is inserted in either a single copy centromeric plasmid or in a multicopy, 2 micron circle-based plasmid.

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Selected References

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

  1. Almer A., Hörz W. Nuclease hypersensitive regions with adjacent positioned nucleosomes mark the gene boundaries of the PHO5/PHO3 locus in yeast. EMBO J. 1986 Oct;5(10):2681–2687. doi: 10.1002/j.1460-2075.1986.tb04551.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Almer A., Rudolph H., Hinnen A., Hörz W. Removal of positioned nucleosomes from the yeast PHO5 promoter upon PHO5 induction releases additional upstream activating DNA elements. EMBO J. 1986 Oct;5(10):2689–2696. doi: 10.1002/j.1460-2075.1986.tb04552.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bergman L. W. A DNA fragment containing the upstream activator sequence determines nucleosome positioning of the transcriptionally repressed PHO5 gene of Saccharomyces cerevisiae. Mol Cell Biol. 1986 Jul;6(7):2298–2304. doi: 10.1128/mcb.6.7.2298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bergman L. W., Kramer R. A. Modulation of chromatin structure associated with derepression of the acid phosphatase gene of Saccharomyces cerevisiae. J Biol Chem. 1983 Jun 10;258(11):7223–7227. [PubMed] [Google Scholar]
  5. Bergman L. W., Stranathan M. C., Preis L. H. Structure of the transcriptionally repressed phosphate-repressible acid phosphatase gene (PHO5) of Saccharomyces cerevisiae. Mol Cell Biol. 1986 Jan;6(1):38–46. doi: 10.1128/mcb.6.1.38. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bloom K. S., Carbon J. Yeast centromere DNA is in a unique and highly ordered structure in chromosomes and small circular minichromosomes. Cell. 1982 Jun;29(2):305–317. doi: 10.1016/0092-8674(82)90147-7. [DOI] [PubMed] [Google Scholar]
  7. Burns D. M., Beacham I. R. A method for the ligation of DNA following isolation from low melting temperature agarose. Anal Biochem. 1983 Nov;135(1):48–51. doi: 10.1016/0003-2697(83)90728-5. [DOI] [PubMed] [Google Scholar]
  8. Carlson M., Botstein D. Two differentially regulated mRNAs with different 5' ends encode secreted with intracellular forms of yeast invertase. Cell. 1982 Jan;28(1):145–154. doi: 10.1016/0092-8674(82)90384-1. [DOI] [PubMed] [Google Scholar]
  9. Carlson M., Osmond B. C., Botstein D. Mutants of yeast defective in sucrose utilization. Genetics. 1981 May;98(1):25–40. doi: 10.1093/genetics/98.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Celenza J. L., Carlson M. A yeast gene that is essential for release from glucose repression encodes a protein kinase. Science. 1986 Sep 12;233(4769):1175–1180. doi: 10.1126/science.3526554. [DOI] [PubMed] [Google Scholar]
  11. Eissenberg J. C., Cartwright I. L., Thomas G. H., Elgin S. C. Selected topics in chromatin structure. Annu Rev Genet. 1985;19:485–536. doi: 10.1146/annurev.ge.19.120185.002413. [DOI] [PubMed] [Google Scholar]
  12. Estruch F., Pérez-Ortín J. E., Franco L. Fractionation of yeast chromatin by micrococcal nuclease digestion. Cell Mol Biol. 1986;32(2):195–199. [PubMed] [Google Scholar]
  13. 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]
  14. Lohr D. A protected region upstream and limited nucleosomal positioning downstream of the transcription initiation region of the yeast 35S ribosomal gene. Biochemistry. 1983 Sep 13;22(19):4527–4534. doi: 10.1021/bi00288a027. [DOI] [PubMed] [Google Scholar]
  15. Lohr D. Chromatin structure differs between coding and upstream flanking sequences of the yeast 35S ribosomal genes. Biochemistry. 1983 Feb 15;22(4):927–934. doi: 10.1021/bi00273a034. [DOI] [PubMed] [Google Scholar]
  16. Lohr D., Hopper J. E. The relationship of regulatory proteins and DNase I hypersensitive sites in the yeast GAL1-10 genes. Nucleic Acids Res. 1985 Dec 9;13(23):8409–8423. doi: 10.1093/nar/13.23.8409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lohr D., Kovacic R. T., Van Holde K. E. Quantitative analysis of the digestion of yeast chromatin by staphylococcal nuclease. Biochemistry. 1977 Feb 8;16(3):463–471. doi: 10.1021/bi00622a020. [DOI] [PubMed] [Google Scholar]
  18. Lohr D. Organization of the GAL1-GAL10 intergenic control region chromatin. Nucleic Acids Res. 1984 Nov 26;12(22):8457–8474. doi: 10.1093/nar/12.22.8457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lohr D. The chromatin structure of an actively expressed, single copy yeast gene. Nucleic Acids Res. 1983 Oct 11;11(19):6755–6773. doi: 10.1093/nar/11.19.6755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lorch Y., LaPointe J. W., Kornberg R. D. Nucleosomes inhibit the initiation of transcription but allow chain elongation with the displacement of histones. Cell. 1987 Apr 24;49(2):203–210. doi: 10.1016/0092-8674(87)90561-7. [DOI] [PubMed] [Google Scholar]
  21. Nedospasov S. A., Georgiev G. P. Non-random cleavage of SV40 DNA in the compact minichromosome and free in solution by micrococcal nuclease. Biochem Biophys Res Commun. 1980 Jan 29;92(2):532–539. doi: 10.1016/0006-291x(80)90366-6. [DOI] [PubMed] [Google Scholar]
  22. Neigeborn L., Carlson M. Genes affecting the regulation of SUC2 gene expression by glucose repression in Saccharomyces cerevisiae. Genetics. 1984 Dec;108(4):845–858. doi: 10.1093/genetics/108.4.845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Neigeborn L., Celenza J. L., Carlson M. SSN20 is an essential gene with mutant alleles that suppress defects in SUC2 transcription in Saccharomyces cerevisiae. Mol Cell Biol. 1987 Feb;7(2):672–678. doi: 10.1128/mcb.7.2.672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pederson D. S., Venkatesan M., Thoma F., Simpson R. T. Isolation of an episomal yeast gene and replication origin as chromatin. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7206–7210. doi: 10.1073/pnas.83.19.7206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Proffitt J. H. DNase I-hypersensitive sites in the galactose gene cluster of Saccharomyces cerevisiae. Mol Cell Biol. 1985 Jun;5(6):1522–1524. doi: 10.1128/mcb.5.6.1522. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pérez-Ortin J. E., Estruch F., Matallana E., Franco L. DNase I sensitivity of the chromatin of the yeast SUC2 gene for invertase. Mol Gen Genet. 1986 Dec;205(3):422–427. doi: 10.1007/BF00338077. [DOI] [PubMed] [Google Scholar]
  27. Pérez-Ortín J. E., Estruch F., Matallana E., Franco L. Sliding-end-labelling. A method to avoid artifacts in nucleosome positioning. FEBS Lett. 1986 Nov 10;208(1):31–33. doi: 10.1016/0014-5793(86)81525-3. [DOI] [PubMed] [Google Scholar]
  28. Renaud J., Ruiz-Carrillo A. Fine analysis of the active H5 gene chromatin of chicken erythroid cells at different stages of differentiation. J Mol Biol. 1986 May 5;189(1):217–226. doi: 10.1016/0022-2836(86)90392-x. [DOI] [PubMed] [Google Scholar]
  29. Ruberti I., Worcel A. Mechanism of chromatin assembly in Xenopus oocytes. J Mol Biol. 1986 Jun 5;189(3):457–476. doi: 10.1016/0022-2836(86)90317-7. [DOI] [PubMed] [Google Scholar]
  30. Sarokin L., Carlson M. Short repeated elements in the upstream regulatory region of the SUC2 gene of Saccharomyces cerevisiae. Mol Cell Biol. 1986 Jul;6(7):2324–2333. doi: 10.1128/mcb.6.7.2324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sarokin L., Carlson M. Upstream region required for regulated expression of the glucose-repressible SUC2 gene of Saccharomyces cerevisiae. Mol Cell Biol. 1984 Dec;4(12):2750–2757. doi: 10.1128/mcb.4.12.2750. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Simpson R. T. Nucleosome positioning in vivo and in vitro. Bioessays. 1986 Apr;4(4):172–176. doi: 10.1002/bies.950040408. [DOI] [PubMed] [Google Scholar]
  33. Sledziewski A., Young E. T. Chromatin conformational changes accompany transcriptional activation of a glucose-repressed gene in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1982 Jan;79(2):253–256. doi: 10.1073/pnas.79.2.253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Szent-Györgyi C., Finkelstein D. B., Garrard W. T. Sharp boundaries demarcate the chromatin structure of a yeast heat-shock gene. J Mol Biol. 1987 Jan 5;193(1):71–80. doi: 10.1016/0022-2836(87)90628-0. [DOI] [PubMed] [Google Scholar]
  35. Thoma F., Bergman L. W., Simpson R. T. Nuclease digestion of circular TRP1ARS1 chromatin reveals positioned nucleosomes separated by nuclease-sensitive regions. J Mol Biol. 1984 Aug 25;177(4):715–733. doi: 10.1016/0022-2836(84)90046-9. [DOI] [PubMed] [Google Scholar]
  36. Thoma F. Protein-DNA interactions and nuclease-sensitive regions determine nucleosome positions on yeast plasmid chromatin. J Mol Biol. 1986 Jul 20;190(2):177–190. doi: 10.1016/0022-2836(86)90291-3. [DOI] [PubMed] [Google Scholar]
  37. Thoma F., Simpson R. T. Local protein-DNA interactions may determine nucleosome positions on yeast plasmids. Nature. 1985 May 16;315(6016):250–252. doi: 10.1038/315250a0. [DOI] [PubMed] [Google Scholar]
  38. Thomas J. O., Furber V. Yeast chromatin structure. FEBS Lett. 1976 Jul 15;66(2):274–280. doi: 10.1016/0014-5793(76)80521-2. [DOI] [PubMed] [Google Scholar]
  39. Wallis J. W., Rykowski M., Grunstein M. Yeast histone H2B containing large amino terminus deletions can function in vivo. Cell. 1983 Dec;35(3 Pt 2):711–719. doi: 10.1016/0092-8674(83)90104-6. [DOI] [PubMed] [Google Scholar]
  40. Wu C., Bingham P. M., Livak K. J., Holmgren R., Elgin S. C. The chromatin structure of specific genes: I. Evidence for higher order domains of defined DNA sequence. Cell. 1979 Apr;16(4):797–806. doi: 10.1016/0092-8674(79)90095-3. [DOI] [PubMed] [Google Scholar]
  41. Wu C. The 5' ends of Drosophila heat shock genes in chromatin are hypersensitive to DNase I. Nature. 1980 Aug 28;286(5776):854–860. doi: 10.1038/286854a0. [DOI] [PubMed] [Google Scholar]

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