Purification of yeast histones competent for nucleosome assembly in vitro

Mariko Fukuma; Yoshiki Hiraoka; Hiroshi Sakurai; Toshio Fukasawa

doi:10.1002/yea.320100305

Purification of yeast histones competent for nucleosome assembly in vitro

Mariko Fukuma, Yoshiki Hiraoka, Hiroshi Sakurai, Toshio Fukasawa

Department of Anatomy

Research output: Contribution to journal › Article › peer-review

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 language	English
Pages (from-to)	319-331
Number of pages	13
Journal	Yeast
Volume	10
Issue number	3
DOIs	https://doi.org/10.1002/yea.320100305
Publication status	Published - 1994 Mar

Keywords

Saccharomyces cerevisiae
histones
nucleosome assembly

ASJC Scopus subject areas

Biotechnology
Bioengineering
Biochemistry
Applied Microbiology and Biotechnology
Genetics

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10.1002/yea.320100305

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title = "Purification of yeast histones competent for nucleosome assembly in vitro",

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.",

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T1 - Purification of yeast histones competent for nucleosome assembly in vitro

AU - Fukuma, Mariko

AU - Hiraoka, Yoshiki

AU - Sakurai, Hiroshi

AU - Fukasawa, Toshio

PY - 1994/3

Y1 - 1994/3

N2 - 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.

AB - 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.

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KW - nucleosome assembly

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DO - 10.1002/yea.320100305

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JO - Yeast

JF - Yeast

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ER -

Purification of yeast histones competent for nucleosome assembly in vitro

Abstract

Keywords

ASJC Scopus subject areas

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