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

    研究成果: Article

    55 引用 (Scopus)

    抄録

    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.

    元の言語English
    ページ(範囲)2208-2218
    ページ数11
    ジャーナルNucleic Acids Research
    36
    発行部数7
    DOI
    出版物ステータスPublished - 2008 4

    Fingerprint

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

    ASJC Scopus subject areas

    • Genetics

    これを引用

    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.

    :: Nucleic Acids Research, 巻 36, 番号 7, 04.2008, p. 2208-2218.

    研究成果: Article

    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. :: Nucleic Acids Research. 2008 ; 巻 36, 番号 7. pp. 2208-2218.
    @article{54098ba06aad4107a788f794581655ab,
    title = "Nucleosome formation with the testis-specific histone H3 variant, H3t, by human nucleosome assembly proteins in vitro",
    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.",
    author = "Hiroaki Tachiwana and Akihisa Osakabe and Hiroshi Kimura and Hitoshi Kurumizaka",
    year = "2008",
    month = "4",
    doi = "10.1093/nar/gkn060",
    language = "English",
    volume = "36",
    pages = "2208--2218",
    journal = "Nucleic Acids Research",
    issn = "0305-1048",
    publisher = "Oxford University Press",
    number = "7",

    }

    TY - JOUR

    T1 - Nucleosome formation with the testis-specific histone H3 variant, H3t, by human nucleosome assembly proteins in vitro

    AU - Tachiwana, Hiroaki

    AU - Osakabe, Akihisa

    AU - Kimura, Hiroshi

    AU - Kurumizaka, Hitoshi

    PY - 2008/4

    Y1 - 2008/4

    N2 - 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.

    AB - 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.

    UR - http://www.scopus.com/inward/record.url?scp=42449097684&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=42449097684&partnerID=8YFLogxK

    U2 - 10.1093/nar/gkn060

    DO - 10.1093/nar/gkn060

    M3 - Article

    VL - 36

    SP - 2208

    EP - 2218

    JO - Nucleic Acids Research

    JF - Nucleic Acids Research

    SN - 0305-1048

    IS - 7

    ER -