Nucleosome formation with the testis-specific histone H3 variant, H3t, by human nucleosome assembly proteins in vitro

Hiroaki Tachiwana, Akihisa Osakabe, Hiroshi Kimura, Hitoshi Kurumizaka

    Research output: Contribution to journalArticle

    55 Citations (Scopus)

    Abstract

    Five non-allelic histone H3 variants, H3.1, H3.2, H3.3, H3t and CENP-A, have been identified in mammals. H3t is robustly expressed in the testis, and thus was assigned as the testis-specific H3 variant. However, recent proteomics and tissue-specific RT-PCR experiments revealed a small amount of H3t expression in somatic cells. In the present study, we purified human H3t as a recombinant protein, and showed that H3t/H4 forms nucleosomes with H2A/H2B by the salt-dialysis method, like the conventional H3.1/H4. We found that H3t/H4 is not efficiently incorporated into the nucleosome by human Nap1 (hNap1), due to its defective H3t/H4 deposition on DNA. In contrast, human Nap2 (hNap2), a paralog of hNap1, promotes nucleosome assembly with H3t/H4. Mutational analyses revealed that the Ala111 residue, which is conserved among H3.1, H3.2 and H3.3, but not in H3t, is the essential residue for the hNap1-mediated nucleosome assembly. These results suggest that H3t may be incorporated into chromatin by a specific chaperone-mediated pathway.

    Original languageEnglish
    Pages (from-to)2208-2218
    Number of pages11
    JournalNucleic Acids Research
    Volume36
    Issue number7
    DOIs
    Publication statusPublished - 2008 Apr

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    Nucleosomes
    Histones
    Testis
    Proteins
    Recombinant Proteins
    Proteomics
    Chromatin
    Dialysis
    Mammals
    Salts
    In Vitro Techniques
    Polymerase Chain Reaction
    DNA

    ASJC Scopus subject areas

    • Genetics

    Cite this

    Nucleosome formation with the testis-specific histone H3 variant, H3t, by human nucleosome assembly proteins in vitro. / Tachiwana, Hiroaki; Osakabe, Akihisa; Kimura, Hiroshi; Kurumizaka, Hitoshi.

    In: Nucleic Acids Research, Vol. 36, No. 7, 04.2008, p. 2208-2218.

    Research output: Contribution to journalArticle

    Tachiwana, Hiroaki ; Osakabe, Akihisa ; Kimura, Hiroshi ; Kurumizaka, Hitoshi. / Nucleosome formation with the testis-specific histone H3 variant, H3t, by human nucleosome assembly proteins in vitro. In: Nucleic Acids Research. 2008 ; Vol. 36, No. 7. pp. 2208-2218.
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