Purification of yeast histones competent for nucleosome assembly in vitro

M. Fukuma, Yoshiki Hiraoka, H. Sakurai, T. Fukasawa

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

We have developed a procedure to purify nucleosomal assembly-competent histones as a mixture of H2A, H2B, H3 and H4 from isolated nuclei of the yeast Saccharomyces cerevisiae with a purity of 70-80%. The mixture contained each of the histone subunits approximately at the equi-molar ratio. Plasmid pBR322 DNA was assembled into nucleosomes with the purified yeast histones in the presence of nucleoplasmin from unfertilized eggs of the frog Xenopus laevis. The efficiency of assembly of yeast histones was comparable to that of core histones purified from HeLa cells. The length of DNA fragment wrapping around a core histone particle and the molar ratio of histone components in an assembled nucleosome particle were estimated to be 150 ± 10 bp long and H2A:H2B:H3:H4=1·0:0·9:0·9:1·0, respectively.

Original languageEnglish
Pages (from-to)319-331
Number of pages13
JournalYeast
Volume10
Issue number3
Publication statusPublished - 1994

Fingerprint

nucleosomes
Nucleosomes
histones
Histones
Yeast
Purification
Yeasts
yeasts
DNA
Nucleoplasmins
Equus
Xenopus laevis
In Vitro Techniques
HeLa Cells
Anura
purity
Ovum
Saccharomyces cerevisiae
frogs
plasmids

Keywords

  • histones
  • nucleosome assembly
  • Saccharomyces cerevisiae

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology
  • Bioengineering
  • Microbiology

Cite this

Fukuma, M., Hiraoka, Y., Sakurai, H., & Fukasawa, T. (1994). Purification of yeast histones competent for nucleosome assembly in vitro. Yeast, 10(3), 319-331.

Purification of yeast histones competent for nucleosome assembly in vitro. / Fukuma, M.; Hiraoka, Yoshiki; Sakurai, H.; Fukasawa, T.

In: Yeast, Vol. 10, No. 3, 1994, p. 319-331.

Research output: Contribution to journalArticle

Fukuma, M, Hiraoka, Y, Sakurai, H & Fukasawa, T 1994, 'Purification of yeast histones competent for nucleosome assembly in vitro', Yeast, vol. 10, no. 3, pp. 319-331.
Fukuma, M. ; Hiraoka, Yoshiki ; Sakurai, H. ; Fukasawa, T. / Purification of yeast histones competent for nucleosome assembly in vitro. In: Yeast. 1994 ; Vol. 10, No. 3. pp. 319-331.
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