Vertebrate Spt2 is a novel nucleolar histone chaperone that assists in ribosomal DNA transcription

Akihisa Osakabe, Hiroaki Tachiwana, Motoki Takaku, Tetsuya Hori, Chikashi Obuse, Hiroshi Kimura, Tatsuo Fukagawa, Hitoshi Kurumizaka

    Research output: Contribution to journalArticle

    11 Citations (Scopus)

    Abstract

    In eukaryotes, transcription occurs in the chromatin context with the assistance of histone-binding proteins, such as chromatin/nucleosome remodeling factors and histone chaperones. However, it is unclear how each remodeling factor or histone chaperone functions in transcription. Here, we identify a novel histone-binding protein, Spt2, in higher eukaryotes. Recombinant human Spt2 binds to histones and DNA, and promotes nucleosome assembly in vitro. Spt2 accumulates in nucleoli and interacts with RNA polymerase I in chicken DT40 cells, suggesting its involvement in ribosomal RNA transcription. Consistently, Spt2-deficient chicken DT40 cells are sensitive to RNA polymerase I inhibitors and exhibit decreased transcription activity, as shown by a transcription run-on assay. Domain analyses of Spt2 revealed that the C-terminal region, containing the region homologous to yeast Spt2, is responsible for histone binding, while the central region is essential for nucleolar localization and DNA binding. Based on these results, we conclude that vertebrate Spt2 is a novel histone chaperone with a separate DNA-binding domain that facilitates ribosomal DNA transcription through chromatin remodeling during transcription.

    Original languageEnglish
    Pages (from-to)1323-1332
    Number of pages10
    JournalJournal of Cell Science
    Volume126
    Issue number6
    DOIs
    Publication statusPublished - 2013 Mar 15

    Fingerprint

    Histone Chaperones
    Ribosomal DNA
    Histones
    Vertebrates
    RNA Polymerase I
    Chromatin Assembly and Disassembly
    Nucleosomes
    Eukaryota
    Chickens
    DNA
    Carrier Proteins
    Ribosomal RNA
    Chromatin
    Yeasts

    Keywords

    • Chromatin
    • Histone chaperone
    • Nucleoli
    • Nucleosome
    • RNA polymerase
    • Transcription

    ASJC Scopus subject areas

    • Cell Biology

    Cite this

    Osakabe, A., Tachiwana, H., Takaku, M., Hori, T., Obuse, C., Kimura, H., ... Kurumizaka, H. (2013). Vertebrate Spt2 is a novel nucleolar histone chaperone that assists in ribosomal DNA transcription. Journal of Cell Science, 126(6), 1323-1332. https://doi.org/10.1242/jcs.112623

    Vertebrate Spt2 is a novel nucleolar histone chaperone that assists in ribosomal DNA transcription. / Osakabe, Akihisa; Tachiwana, Hiroaki; Takaku, Motoki; Hori, Tetsuya; Obuse, Chikashi; Kimura, Hiroshi; Fukagawa, Tatsuo; Kurumizaka, Hitoshi.

    In: Journal of Cell Science, Vol. 126, No. 6, 15.03.2013, p. 1323-1332.

    Research output: Contribution to journalArticle

    Osakabe, A, Tachiwana, H, Takaku, M, Hori, T, Obuse, C, Kimura, H, Fukagawa, T & Kurumizaka, H 2013, 'Vertebrate Spt2 is a novel nucleolar histone chaperone that assists in ribosomal DNA transcription', Journal of Cell Science, vol. 126, no. 6, pp. 1323-1332. https://doi.org/10.1242/jcs.112623
    Osakabe, Akihisa ; Tachiwana, Hiroaki ; Takaku, Motoki ; Hori, Tetsuya ; Obuse, Chikashi ; Kimura, Hiroshi ; Fukagawa, Tatsuo ; Kurumizaka, Hitoshi. / Vertebrate Spt2 is a novel nucleolar histone chaperone that assists in ribosomal DNA transcription. In: Journal of Cell Science. 2013 ; Vol. 126, No. 6. pp. 1323-1332.
    @article{b5161e14e458491ca3b791bfd4398622,
    title = "Vertebrate Spt2 is a novel nucleolar histone chaperone that assists in ribosomal DNA transcription",
    abstract = "In eukaryotes, transcription occurs in the chromatin context with the assistance of histone-binding proteins, such as chromatin/nucleosome remodeling factors and histone chaperones. However, it is unclear how each remodeling factor or histone chaperone functions in transcription. Here, we identify a novel histone-binding protein, Spt2, in higher eukaryotes. Recombinant human Spt2 binds to histones and DNA, and promotes nucleosome assembly in vitro. Spt2 accumulates in nucleoli and interacts with RNA polymerase I in chicken DT40 cells, suggesting its involvement in ribosomal RNA transcription. Consistently, Spt2-deficient chicken DT40 cells are sensitive to RNA polymerase I inhibitors and exhibit decreased transcription activity, as shown by a transcription run-on assay. Domain analyses of Spt2 revealed that the C-terminal region, containing the region homologous to yeast Spt2, is responsible for histone binding, while the central region is essential for nucleolar localization and DNA binding. Based on these results, we conclude that vertebrate Spt2 is a novel histone chaperone with a separate DNA-binding domain that facilitates ribosomal DNA transcription through chromatin remodeling during transcription.",
    keywords = "Chromatin, Histone chaperone, Nucleoli, Nucleosome, RNA polymerase, Transcription",
    author = "Akihisa Osakabe and Hiroaki Tachiwana and Motoki Takaku and Tetsuya Hori and Chikashi Obuse and Hiroshi Kimura and Tatsuo Fukagawa and Hitoshi Kurumizaka",
    year = "2013",
    month = "3",
    day = "15",
    doi = "10.1242/jcs.112623",
    language = "English",
    volume = "126",
    pages = "1323--1332",
    journal = "Journal of Cell Science",
    issn = "0021-9533",
    publisher = "Company of Biologists Ltd",
    number = "6",

    }

    TY - JOUR

    T1 - Vertebrate Spt2 is a novel nucleolar histone chaperone that assists in ribosomal DNA transcription

    AU - Osakabe, Akihisa

    AU - Tachiwana, Hiroaki

    AU - Takaku, Motoki

    AU - Hori, Tetsuya

    AU - Obuse, Chikashi

    AU - Kimura, Hiroshi

    AU - Fukagawa, Tatsuo

    AU - Kurumizaka, Hitoshi

    PY - 2013/3/15

    Y1 - 2013/3/15

    N2 - In eukaryotes, transcription occurs in the chromatin context with the assistance of histone-binding proteins, such as chromatin/nucleosome remodeling factors and histone chaperones. However, it is unclear how each remodeling factor or histone chaperone functions in transcription. Here, we identify a novel histone-binding protein, Spt2, in higher eukaryotes. Recombinant human Spt2 binds to histones and DNA, and promotes nucleosome assembly in vitro. Spt2 accumulates in nucleoli and interacts with RNA polymerase I in chicken DT40 cells, suggesting its involvement in ribosomal RNA transcription. Consistently, Spt2-deficient chicken DT40 cells are sensitive to RNA polymerase I inhibitors and exhibit decreased transcription activity, as shown by a transcription run-on assay. Domain analyses of Spt2 revealed that the C-terminal region, containing the region homologous to yeast Spt2, is responsible for histone binding, while the central region is essential for nucleolar localization and DNA binding. Based on these results, we conclude that vertebrate Spt2 is a novel histone chaperone with a separate DNA-binding domain that facilitates ribosomal DNA transcription through chromatin remodeling during transcription.

    AB - In eukaryotes, transcription occurs in the chromatin context with the assistance of histone-binding proteins, such as chromatin/nucleosome remodeling factors and histone chaperones. However, it is unclear how each remodeling factor or histone chaperone functions in transcription. Here, we identify a novel histone-binding protein, Spt2, in higher eukaryotes. Recombinant human Spt2 binds to histones and DNA, and promotes nucleosome assembly in vitro. Spt2 accumulates in nucleoli and interacts with RNA polymerase I in chicken DT40 cells, suggesting its involvement in ribosomal RNA transcription. Consistently, Spt2-deficient chicken DT40 cells are sensitive to RNA polymerase I inhibitors and exhibit decreased transcription activity, as shown by a transcription run-on assay. Domain analyses of Spt2 revealed that the C-terminal region, containing the region homologous to yeast Spt2, is responsible for histone binding, while the central region is essential for nucleolar localization and DNA binding. Based on these results, we conclude that vertebrate Spt2 is a novel histone chaperone with a separate DNA-binding domain that facilitates ribosomal DNA transcription through chromatin remodeling during transcription.

    KW - Chromatin

    KW - Histone chaperone

    KW - Nucleoli

    KW - Nucleosome

    KW - RNA polymerase

    KW - Transcription

    UR - http://www.scopus.com/inward/record.url?scp=84877917618&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=84877917618&partnerID=8YFLogxK

    U2 - 10.1242/jcs.112623

    DO - 10.1242/jcs.112623

    M3 - Article

    VL - 126

    SP - 1323

    EP - 1332

    JO - Journal of Cell Science

    JF - Journal of Cell Science

    SN - 0021-9533

    IS - 6

    ER -