Quaternary structure of the human Cdt1-Geminin complex regulates DNA replication licensing

V. De Marco, P. J. Gillespie, A. Li, N. Karantzelis, E. Christodoulou, R. Klompmaker, S. van Gerwen, A. Fish, M. V. Petoukhov, M. S. Iliou, Z. Lygerou, R. H. Medema, J.J. Blow, D. I. Svergun, S. Taraviras, A. Perrakis

    Research output: Contribution to journalArticle

    48 Citations (Scopus)

    Abstract

    All organisms need to ensure that no DNA segments are rereplicated in a single cell cycle. Eukaryotes achieve this through a process called origin licensing, which involves tight spatiotemporal control of the assembly of prereplicative complexes (pre-RCs) onto chromatin. Cdt1 is a key component and crucial regulator of pre-RC assembly. In higher eukaryotes, timely inhibition of Cdt1 by Geminin is essential to prevent DNA rereplication. Here, we address the mechanism of DNA licensing inhibition by Geminin, by combining X-ray crystallography, small-angle X-ray scattering, and functional studies in Xenopus and mammalian cells. Our findings show that the Cdt1: Geminin complex can exist in two distinct forms, a "permissive" heterotrimer and an "inhibitory" heterohexamer. Specific Cdt1 residues, buried in the heterohexamer, are important for licensing. We postulate that the transition between the heterotrimer and the heterohexamer represents a molecular switch between licensing-competent and licensing-defective states.

    Original languageEnglish
    Pages (from-to)19807-19812
    Number of pages6
    JournalProceedings of the National Academy of Sciences of the United States of America
    Volume106
    Issue number47
    DOIs
    Publication statusPublished - 2009

    Keywords

    • solution structure
    • X-ray structure
    • pre-RC
    • cell cycle
    • XENOPUS EGG EXTRACTS
    • CELL-CYCLE
    • RE-REPLICATION
    • FACTOR CDT1
    • PREVENTS REREPLICATION
    • NUCLEAR IMPORT
    • S-PHASE
    • GEMININ
    • INHIBITION
    • DESTRUCTION

    Cite this

    De Marco, V. ; Gillespie, P. J. ; Li, A. ; Karantzelis, N. ; Christodoulou, E. ; Klompmaker, R. ; van Gerwen, S. ; Fish, A. ; Petoukhov, M. V. ; Iliou, M. S. ; Lygerou, Z. ; Medema, R. H. ; Blow, J.J. ; Svergun, D. I. ; Taraviras, S. ; Perrakis, A. / Quaternary structure of the human Cdt1-Geminin complex regulates DNA replication licensing. In: Proceedings of the National Academy of Sciences of the United States of America. 2009 ; Vol. 106, No. 47. pp. 19807-19812.
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    title = "Quaternary structure of the human Cdt1-Geminin complex regulates DNA replication licensing",
    abstract = "All organisms need to ensure that no DNA segments are rereplicated in a single cell cycle. Eukaryotes achieve this through a process called origin licensing, which involves tight spatiotemporal control of the assembly of prereplicative complexes (pre-RCs) onto chromatin. Cdt1 is a key component and crucial regulator of pre-RC assembly. In higher eukaryotes, timely inhibition of Cdt1 by Geminin is essential to prevent DNA rereplication. Here, we address the mechanism of DNA licensing inhibition by Geminin, by combining X-ray crystallography, small-angle X-ray scattering, and functional studies in Xenopus and mammalian cells. Our findings show that the Cdt1: Geminin complex can exist in two distinct forms, a {"}permissive{"} heterotrimer and an {"}inhibitory{"} heterohexamer. Specific Cdt1 residues, buried in the heterohexamer, are important for licensing. We postulate that the transition between the heterotrimer and the heterohexamer represents a molecular switch between licensing-competent and licensing-defective states.",
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    author = "{De Marco}, V. and Gillespie, {P. J.} and A. Li and N. Karantzelis and E. Christodoulou and R. Klompmaker and {van Gerwen}, S. and A. Fish and Petoukhov, {M. V.} and Iliou, {M. S.} and Z. Lygerou and Medema, {R. H.} and J.J. Blow and Svergun, {D. I.} and S. Taraviras and A. Perrakis",
    year = "2009",
    doi = "10.1073/pnas.0905281106",
    language = "English",
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    pages = "19807--19812",
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    De Marco, V, Gillespie, PJ, Li, A, Karantzelis, N, Christodoulou, E, Klompmaker, R, van Gerwen, S, Fish, A, Petoukhov, MV, Iliou, MS, Lygerou, Z, Medema, RH, Blow, JJ, Svergun, DI, Taraviras, S & Perrakis, A 2009, 'Quaternary structure of the human Cdt1-Geminin complex regulates DNA replication licensing', Proceedings of the National Academy of Sciences of the United States of America, vol. 106, no. 47, pp. 19807-19812. https://doi.org/10.1073/pnas.0905281106

    Quaternary structure of the human Cdt1-Geminin complex regulates DNA replication licensing. / De Marco, V.; Gillespie, P. J. ; Li, A.; Karantzelis, N.; Christodoulou, E.; Klompmaker, R.; van Gerwen, S.; Fish, A.; Petoukhov, M. V.; Iliou, M. S.; Lygerou, Z.; Medema, R. H.; Blow, J.J.; Svergun, D. I.; Taraviras, S.; Perrakis, A.

    In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 106, No. 47, 2009, p. 19807-19812.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Quaternary structure of the human Cdt1-Geminin complex regulates DNA replication licensing

    AU - De Marco, V.

    AU - Gillespie, P. J.

    AU - Li, A.

    AU - Karantzelis, N.

    AU - Christodoulou, E.

    AU - Klompmaker, R.

    AU - van Gerwen, S.

    AU - Fish, A.

    AU - Petoukhov, M. V.

    AU - Iliou, M. S.

    AU - Lygerou, Z.

    AU - Medema, R. H.

    AU - Blow, J.J.

    AU - Svergun, D. I.

    AU - Taraviras, S.

    AU - Perrakis, A.

    PY - 2009

    Y1 - 2009

    N2 - All organisms need to ensure that no DNA segments are rereplicated in a single cell cycle. Eukaryotes achieve this through a process called origin licensing, which involves tight spatiotemporal control of the assembly of prereplicative complexes (pre-RCs) onto chromatin. Cdt1 is a key component and crucial regulator of pre-RC assembly. In higher eukaryotes, timely inhibition of Cdt1 by Geminin is essential to prevent DNA rereplication. Here, we address the mechanism of DNA licensing inhibition by Geminin, by combining X-ray crystallography, small-angle X-ray scattering, and functional studies in Xenopus and mammalian cells. Our findings show that the Cdt1: Geminin complex can exist in two distinct forms, a "permissive" heterotrimer and an "inhibitory" heterohexamer. Specific Cdt1 residues, buried in the heterohexamer, are important for licensing. We postulate that the transition between the heterotrimer and the heterohexamer represents a molecular switch between licensing-competent and licensing-defective states.

    AB - All organisms need to ensure that no DNA segments are rereplicated in a single cell cycle. Eukaryotes achieve this through a process called origin licensing, which involves tight spatiotemporal control of the assembly of prereplicative complexes (pre-RCs) onto chromatin. Cdt1 is a key component and crucial regulator of pre-RC assembly. In higher eukaryotes, timely inhibition of Cdt1 by Geminin is essential to prevent DNA rereplication. Here, we address the mechanism of DNA licensing inhibition by Geminin, by combining X-ray crystallography, small-angle X-ray scattering, and functional studies in Xenopus and mammalian cells. Our findings show that the Cdt1: Geminin complex can exist in two distinct forms, a "permissive" heterotrimer and an "inhibitory" heterohexamer. Specific Cdt1 residues, buried in the heterohexamer, are important for licensing. We postulate that the transition between the heterotrimer and the heterohexamer represents a molecular switch between licensing-competent and licensing-defective states.

    KW - solution structure

    KW - X-ray structure

    KW - pre-RC

    KW - cell cycle

    KW - XENOPUS EGG EXTRACTS

    KW - CELL-CYCLE

    KW - RE-REPLICATION

    KW - FACTOR CDT1

    KW - PREVENTS REREPLICATION

    KW - NUCLEAR IMPORT

    KW - S-PHASE

    KW - GEMININ

    KW - INHIBITION

    KW - DESTRUCTION

    U2 - 10.1073/pnas.0905281106

    DO - 10.1073/pnas.0905281106

    M3 - Article

    C2 - 19906994

    VL - 106

    SP - 19807

    EP - 19812

    JO - Proceedings of the National Academy of Sciences

    JF - Proceedings of the National Academy of Sciences

    SN - 0027-8424

    IS - 47

    ER -