HSP90 and Its R2TP/Prefoldin-like Cochaperone Are Involved in the Cytoplasmic Assembly of RNA Polymerase II

Severine Boulon, Berengere Pradet-Balade, Celine Verheggen, Dorothee Molle, Stephanie Boireau, Marya Georgieva, Karim Azzag, Marie-Cecile Robert, Yasmeen Ahmad, Henry Neel, Angus I. Lamond, Edouard Bertrand

    Research output: Contribution to journalArticle

    143 Citations (Scopus)

    Abstract

    RNA polymerases are key multisubunit cellular enzymes. Microscopy studies indicated that RNA polymerase I assembles near its promoter. However, the mechanism by which RNA polymerase II is assembled from its 12 subunits remains unclear. We show here that RNA polymerase II subunits Rpb1 and Rpb3 accumulate in the cytoplasm when assembly is prevented and that nuclear import of Rpb1 requires the presence of all subunits. Using MS-based quantitative proteomics, we characterized assembly intermediates. These included a cytoplasmic complex containing subunits Rpb1 and Rpb8 associated with the HSP90 cochaperone hSpagh (RPAP3) and the R2TP/Prefoldin-like complex. Remarkably, HSP90 activity stabilized incompletely assembled Rpb1 in the cytoplasm. Our data indicate that RNA polymerase II is built in the cytoplasm and reveal quality-control mechanisms that link HSP90 to the nuclear import of fully assembled enzymes. hSpagh also bound the free RPA194 subunit of RNA polymerase I, suggesting a general role in assembling RNA polymerases.

    Original languageEnglish
    Pages (from-to)912-924
    Number of pages13
    JournalMolecular Cell
    Volume39
    Issue number6
    DOIs
    Publication statusPublished - 24 Sep 2010

    Keywords

    • HUMAN TRANSCRIPTION MACHINERY
    • PROTEIN-INTERACTION NETWORK
    • IN-VIVO
    • QUANTITATIVE PROTEOMICS
    • GENE-EXPRESSION
    • LIVING CELLS
    • REAL-TIME
    • DYNAMICS
    • CHAPERONE
    • RECEPTOR

    Cite this

    Boulon, S., Pradet-Balade, B., Verheggen, C., Molle, D., Boireau, S., Georgieva, M., ... Bertrand, E. (2010). HSP90 and Its R2TP/Prefoldin-like Cochaperone Are Involved in the Cytoplasmic Assembly of RNA Polymerase II. Molecular Cell, 39(6), 912-924. https://doi.org/10.1016/j.molcel.2010.08.023
    Boulon, Severine ; Pradet-Balade, Berengere ; Verheggen, Celine ; Molle, Dorothee ; Boireau, Stephanie ; Georgieva, Marya ; Azzag, Karim ; Robert, Marie-Cecile ; Ahmad, Yasmeen ; Neel, Henry ; Lamond, Angus I. ; Bertrand, Edouard. / HSP90 and Its R2TP/Prefoldin-like Cochaperone Are Involved in the Cytoplasmic Assembly of RNA Polymerase II. In: Molecular Cell. 2010 ; Vol. 39, No. 6. pp. 912-924.
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    title = "HSP90 and Its R2TP/Prefoldin-like Cochaperone Are Involved in the Cytoplasmic Assembly of RNA Polymerase II",
    abstract = "RNA polymerases are key multisubunit cellular enzymes. Microscopy studies indicated that RNA polymerase I assembles near its promoter. However, the mechanism by which RNA polymerase II is assembled from its 12 subunits remains unclear. We show here that RNA polymerase II subunits Rpb1 and Rpb3 accumulate in the cytoplasm when assembly is prevented and that nuclear import of Rpb1 requires the presence of all subunits. Using MS-based quantitative proteomics, we characterized assembly intermediates. These included a cytoplasmic complex containing subunits Rpb1 and Rpb8 associated with the HSP90 cochaperone hSpagh (RPAP3) and the R2TP/Prefoldin-like complex. Remarkably, HSP90 activity stabilized incompletely assembled Rpb1 in the cytoplasm. Our data indicate that RNA polymerase II is built in the cytoplasm and reveal quality-control mechanisms that link HSP90 to the nuclear import of fully assembled enzymes. hSpagh also bound the free RPA194 subunit of RNA polymerase I, suggesting a general role in assembling RNA polymerases.",
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    author = "Severine Boulon and Berengere Pradet-Balade and Celine Verheggen and Dorothee Molle and Stephanie Boireau and Marya Georgieva and Karim Azzag and Marie-Cecile Robert and Yasmeen Ahmad and Henry Neel and Lamond, {Angus I.} and Edouard Bertrand",
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    Boulon, S, Pradet-Balade, B, Verheggen, C, Molle, D, Boireau, S, Georgieva, M, Azzag, K, Robert, M-C, Ahmad, Y, Neel, H, Lamond, AI & Bertrand, E 2010, 'HSP90 and Its R2TP/Prefoldin-like Cochaperone Are Involved in the Cytoplasmic Assembly of RNA Polymerase II', Molecular Cell, vol. 39, no. 6, pp. 912-924. https://doi.org/10.1016/j.molcel.2010.08.023

    HSP90 and Its R2TP/Prefoldin-like Cochaperone Are Involved in the Cytoplasmic Assembly of RNA Polymerase II. / Boulon, Severine; Pradet-Balade, Berengere; Verheggen, Celine; Molle, Dorothee; Boireau, Stephanie; Georgieva, Marya; Azzag, Karim; Robert, Marie-Cecile; Ahmad, Yasmeen; Neel, Henry; Lamond, Angus I.; Bertrand, Edouard.

    In: Molecular Cell, Vol. 39, No. 6, 24.09.2010, p. 912-924.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - HSP90 and Its R2TP/Prefoldin-like Cochaperone Are Involved in the Cytoplasmic Assembly of RNA Polymerase II

    AU - Boulon, Severine

    AU - Pradet-Balade, Berengere

    AU - Verheggen, Celine

    AU - Molle, Dorothee

    AU - Boireau, Stephanie

    AU - Georgieva, Marya

    AU - Azzag, Karim

    AU - Robert, Marie-Cecile

    AU - Ahmad, Yasmeen

    AU - Neel, Henry

    AU - Lamond, Angus I.

    AU - Bertrand, Edouard

    PY - 2010/9/24

    Y1 - 2010/9/24

    N2 - RNA polymerases are key multisubunit cellular enzymes. Microscopy studies indicated that RNA polymerase I assembles near its promoter. However, the mechanism by which RNA polymerase II is assembled from its 12 subunits remains unclear. We show here that RNA polymerase II subunits Rpb1 and Rpb3 accumulate in the cytoplasm when assembly is prevented and that nuclear import of Rpb1 requires the presence of all subunits. Using MS-based quantitative proteomics, we characterized assembly intermediates. These included a cytoplasmic complex containing subunits Rpb1 and Rpb8 associated with the HSP90 cochaperone hSpagh (RPAP3) and the R2TP/Prefoldin-like complex. Remarkably, HSP90 activity stabilized incompletely assembled Rpb1 in the cytoplasm. Our data indicate that RNA polymerase II is built in the cytoplasm and reveal quality-control mechanisms that link HSP90 to the nuclear import of fully assembled enzymes. hSpagh also bound the free RPA194 subunit of RNA polymerase I, suggesting a general role in assembling RNA polymerases.

    AB - RNA polymerases are key multisubunit cellular enzymes. Microscopy studies indicated that RNA polymerase I assembles near its promoter. However, the mechanism by which RNA polymerase II is assembled from its 12 subunits remains unclear. We show here that RNA polymerase II subunits Rpb1 and Rpb3 accumulate in the cytoplasm when assembly is prevented and that nuclear import of Rpb1 requires the presence of all subunits. Using MS-based quantitative proteomics, we characterized assembly intermediates. These included a cytoplasmic complex containing subunits Rpb1 and Rpb8 associated with the HSP90 cochaperone hSpagh (RPAP3) and the R2TP/Prefoldin-like complex. Remarkably, HSP90 activity stabilized incompletely assembled Rpb1 in the cytoplasm. Our data indicate that RNA polymerase II is built in the cytoplasm and reveal quality-control mechanisms that link HSP90 to the nuclear import of fully assembled enzymes. hSpagh also bound the free RPA194 subunit of RNA polymerase I, suggesting a general role in assembling RNA polymerases.

    KW - HUMAN TRANSCRIPTION MACHINERY

    KW - PROTEIN-INTERACTION NETWORK

    KW - IN-VIVO

    KW - QUANTITATIVE PROTEOMICS

    KW - GENE-EXPRESSION

    KW - LIVING CELLS

    KW - REAL-TIME

    KW - DYNAMICS

    KW - CHAPERONE

    KW - RECEPTOR

    U2 - 10.1016/j.molcel.2010.08.023

    DO - 10.1016/j.molcel.2010.08.023

    M3 - Article

    VL - 39

    SP - 912

    EP - 924

    JO - Molecular Cell

    JF - Molecular Cell

    SN - 1097-2765

    IS - 6

    ER -

    Boulon S, Pradet-Balade B, Verheggen C, Molle D, Boireau S, Georgieva M et al. HSP90 and Its R2TP/Prefoldin-like Cochaperone Are Involved in the Cytoplasmic Assembly of RNA Polymerase II. Molecular Cell. 2010 Sep 24;39(6):912-924. https://doi.org/10.1016/j.molcel.2010.08.023