Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1997 May 1;25(9):1872–1873. doi: 10.1093/nar/25.9.1872

Rapid isolation of yeast plasmids as native chromatin.

C Martinez-Campa 1, N A Kent 1, J Mellor 1
PMCID: PMC146668  PMID: 9108177

Abstract

Many regions of chromatin are subject to dynamic changes. We have developed a rapid method for isolation of small chromatin templates from yeast which will facilitate biochemical analysis of chromatin composition. Using the PHO5 promoter we show that templates prepared from cells grown in inducing or repressing conditions show native chromatin structures. This method may be widely applicable as the chromatin structures at a centromere, at ARS1 and at part of the lacZ region on two other plasmids are preserved after chromatin isolation.

Full Text

The Full Text of this article is available as a PDF (70.7 KB).

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. Christianson T. W., Sikorski R. S., Dante M., Shero J. H., Hieter P. Multifunctional yeast high-copy-number shuttle vectors. Gene. 1992 Jan 2;110(1):119–122. doi: 10.1016/0378-1119(92)90454-w. [DOI] [PubMed] [Google Scholar]
  4. Fedor M. J., Lue N. F., Kornberg R. D. Statistical positioning of nucleosomes by specific protein-binding to an upstream activating sequence in yeast. J Mol Biol. 1988 Nov 5;204(1):109–127. doi: 10.1016/0022-2836(88)90603-1. [DOI] [PubMed] [Google Scholar]
  5. Kent N. A., Bird L. E., Mellor J. Chromatin analysis in yeast using NP-40 permeabilised sphaeroplasts. Nucleic Acids Res. 1993 Sep 25;21(19):4653–4654. doi: 10.1093/nar/21.19.4653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kent N. A., Mellor J. Chromatin structure snap-shots: rapid nuclease digestion of chromatin in yeast. Nucleic Acids Res. 1995 Sep 25;23(18):3786–3787. doi: 10.1093/nar/23.18.3786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Steger D. J., Workman J. L. Remodeling chromatin structures for transcription: what happens to the histones? Bioessays. 1996 Nov;18(11):875–884. doi: 10.1002/bies.950181106. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Venter U., Svaren J., Schmitz J., Schmid A., Hörz W. A nucleosome precludes binding of the transcription factor Pho4 in vivo to a critical target site in the PHO5 promoter. EMBO J. 1994 Oct 17;13(20):4848–4855. doi: 10.1002/j.1460-2075.1994.tb06811.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES