In vitro reconstitution and biochemical analyses of the Schizosaccharomyces pombe nucleosome

Masako Koyama, Wataru Nagakura, Hiroki Tanaka, Tomoya Kujirai, Yuji Chikashige, Tokuko Haraguchi, Yasushi Hiraoka, Hitoshi Kurumizaka

    Research output: Contribution to journalArticle

    2 Citations (Scopus)

    Abstract

    Schizosaccharomyces pombe, which has a small genome but shares many physiological functions with higher eukaryotes, is a useful single-cell, model eukaryotic organism. In particular, many features concerning chromatin structure and dynamics, including heterochromatin, centromeres, telomeres, and DNA replication origins, are well conserved between S. pombe and higher eukaryotes. However, the S. pombe nucleosome, the fundamental structural unit of chromatin, has not been reconstituted in vitro. In the present study, we established the method to purify S. pombe histones H2A, H2B, H3, and H4, and successfully reconstituted the S. pombe nucleosome in vitro. Our thermal stability assay and micrococcal nuclease treatment assay revealed that the S. pombe nucleosome is markedly unstable and its DNA ends are quite accessible, as compared to the canonical human nucleosome. These findings are important to understand the mechanisms of epigenetic genomic DNA regulation in fission yeast.

    Original languageEnglish
    Pages (from-to)896-901
    Number of pages6
    JournalBiochemical and Biophysical Research Communications
    Volume482
    Issue number4
    DOIs
    Publication statusPublished - 2017 Jan 22

    Fingerprint

    Schizosaccharomyces
    Nucleosomes
    Chromatin
    Assays
    DNA
    Micrococcal Nuclease
    Replication Origin
    Eukaryota
    Histones
    Yeast
    Thermodynamic stability
    Genes
    Heterochromatin
    Centromere
    Telomere
    Eukaryotic Cells
    In Vitro Techniques
    DNA Replication
    Epigenomics
    Hot Temperature

    Keywords

    • Histones
    • Micrococcal nuclease
    • Nucleosome
    • Reconstitution
    • Schizosaccharomyces pombe
    • Thermal stability

    ASJC Scopus subject areas

    • Biophysics
    • Biochemistry
    • Molecular Biology
    • Cell Biology

    Cite this

    Koyama, M., Nagakura, W., Tanaka, H., Kujirai, T., Chikashige, Y., Haraguchi, T., ... Kurumizaka, H. (2017). In vitro reconstitution and biochemical analyses of the Schizosaccharomyces pombe nucleosome. Biochemical and Biophysical Research Communications, 482(4), 896-901. https://doi.org/10.1016/j.bbrc.2016.11.130

    In vitro reconstitution and biochemical analyses of the Schizosaccharomyces pombe nucleosome. / Koyama, Masako; Nagakura, Wataru; Tanaka, Hiroki; Kujirai, Tomoya; Chikashige, Yuji; Haraguchi, Tokuko; Hiraoka, Yasushi; Kurumizaka, Hitoshi.

    In: Biochemical and Biophysical Research Communications, Vol. 482, No. 4, 22.01.2017, p. 896-901.

    Research output: Contribution to journalArticle

    Koyama, M, Nagakura, W, Tanaka, H, Kujirai, T, Chikashige, Y, Haraguchi, T, Hiraoka, Y & Kurumizaka, H 2017, 'In vitro reconstitution and biochemical analyses of the Schizosaccharomyces pombe nucleosome', Biochemical and Biophysical Research Communications, vol. 482, no. 4, pp. 896-901. https://doi.org/10.1016/j.bbrc.2016.11.130
    Koyama, Masako ; Nagakura, Wataru ; Tanaka, Hiroki ; Kujirai, Tomoya ; Chikashige, Yuji ; Haraguchi, Tokuko ; Hiraoka, Yasushi ; Kurumizaka, Hitoshi. / In vitro reconstitution and biochemical analyses of the Schizosaccharomyces pombe nucleosome. In: Biochemical and Biophysical Research Communications. 2017 ; Vol. 482, No. 4. pp. 896-901.
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