Unique binding interactions among Ubc9, SUMO and RanBP2 reveal a mechanism for SUMO paralog selection

Michael H. Tatham, Suhkmann Kim, Ellis Jaffray, Jing Song, Yuan Chen, Ronald T. Hay

    Research output: Contribution to journalArticle

    97 Citations (Scopus)

    Abstract

    The conjugation of small ubiquitin-like modifiers SUMO-1, SUMO-2 and SUMO-3 onto target proteins requires the concerted action of the specific E1-activating enzyme SAE1/SAE2, the E2-conjugating enzyme Ubc9, and an E3-like SUMO ligase. NMR chemical shift perturbation was used to identify the surface of Ubc9 that interacts with the SUMO ligase RanBP2. Unlike known ubiquitin E2-E3 interactions, RanBP2 binds to the -sheet of Ubc9. Mutational disruption of Ubc9-RanBP2 binding affected SUMO-2 but not SUMO-1 conjugation to Sp100 and to a newly identified RanBP2 substrate, PML. RanBP2 contains a binding site specific for SUMO-1 but not SUMO-2, indicating that a Ubc9–SUMO-1 thioester could be recruited to RanBP2 via SUMO-1 in the absence of strong binding between Ubc9 and RanBP2. Thus we show that E2-E3 interactions are not conserved across the ubiquitin-like protein superfamily and identify a RanBP2-dependent mechanism for SUMO paralog–specific conjugation.
    Original languageEnglish
    Pages (from-to)67-74
    Number of pages8
    JournalNature Structural & Molecular Biology
    Volume12
    Issue number1
    DOIs
    Publication statusPublished - Jan 2005

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    Sumoylation
    Ligases
    Ubiquitin
    Ubiquitins
    Enzymes
    Binding Sites
    Proteins

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    @article{ebb2b7b8b4f14c70ae73f1a45ce28fec,
    title = "Unique binding interactions among Ubc9, SUMO and RanBP2 reveal a mechanism for SUMO paralog selection",
    abstract = "The conjugation of small ubiquitin-like modifiers SUMO-1, SUMO-2 and SUMO-3 onto target proteins requires the concerted action of the specific E1-activating enzyme SAE1/SAE2, the E2-conjugating enzyme Ubc9, and an E3-like SUMO ligase. NMR chemical shift perturbation was used to identify the surface of Ubc9 that interacts with the SUMO ligase RanBP2. Unlike known ubiquitin E2-E3 interactions, RanBP2 binds to the -sheet of Ubc9. Mutational disruption of Ubc9-RanBP2 binding affected SUMO-2 but not SUMO-1 conjugation to Sp100 and to a newly identified RanBP2 substrate, PML. RanBP2 contains a binding site specific for SUMO-1 but not SUMO-2, indicating that a Ubc9–SUMO-1 thioester could be recruited to RanBP2 via SUMO-1 in the absence of strong binding between Ubc9 and RanBP2. Thus we show that E2-E3 interactions are not conserved across the ubiquitin-like protein superfamily and identify a RanBP2-dependent mechanism for SUMO paralog–specific conjugation.",
    author = "Tatham, {Michael H.} and Suhkmann Kim and Ellis Jaffray and Jing Song and Yuan Chen and Hay, {Ronald T.}",
    note = "dc.publisher: Nature Publishing Group dc.description.sponsorship: UK Medical Research Council Association for International Cancer Research",
    year = "2005",
    month = "1",
    doi = "10.1038/nsmb878",
    language = "English",
    volume = "12",
    pages = "67--74",
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    Unique binding interactions among Ubc9, SUMO and RanBP2 reveal a mechanism for SUMO paralog selection. / Tatham, Michael H.; Kim, Suhkmann; Jaffray, Ellis; Song, Jing; Chen, Yuan; Hay, Ronald T.

    In: Nature Structural & Molecular Biology, Vol. 12, No. 1, 01.2005, p. 67-74.

    Research output: Contribution to journalArticle

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    AU - Kim, Suhkmann

    AU - Jaffray, Ellis

    AU - Song, Jing

    AU - Chen, Yuan

    AU - Hay, Ronald T.

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    AB - The conjugation of small ubiquitin-like modifiers SUMO-1, SUMO-2 and SUMO-3 onto target proteins requires the concerted action of the specific E1-activating enzyme SAE1/SAE2, the E2-conjugating enzyme Ubc9, and an E3-like SUMO ligase. NMR chemical shift perturbation was used to identify the surface of Ubc9 that interacts with the SUMO ligase RanBP2. Unlike known ubiquitin E2-E3 interactions, RanBP2 binds to the -sheet of Ubc9. Mutational disruption of Ubc9-RanBP2 binding affected SUMO-2 but not SUMO-1 conjugation to Sp100 and to a newly identified RanBP2 substrate, PML. RanBP2 contains a binding site specific for SUMO-1 but not SUMO-2, indicating that a Ubc9–SUMO-1 thioester could be recruited to RanBP2 via SUMO-1 in the absence of strong binding between Ubc9 and RanBP2. Thus we show that E2-E3 interactions are not conserved across the ubiquitin-like protein superfamily and identify a RanBP2-dependent mechanism for SUMO paralog–specific conjugation.

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