A dual E3 mechanism for Rub1 Ligation to Cdc53

Daniel C. Scott, Julie K. Monda, Christy R. R. Grace, David M. Duda, Richard W. Kriwacki, Thimo Kurz, Brenda A. Schulman

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    72 Citations (Scopus)

    Abstract

    In ubiquitin-like protein (UBL) cascades, a thioester-linked E2 similar to UBL complex typically interacts with an E3 enzyme for UBL transfer to the target. Here we demonstrate a variant mechanism, whereby the E2 Ubc12 functions with two E3s, Hrt1 and Dcn1, for ligation of the UBL Rub1 to Cdc53's WHB subdomain. Hrt1 functions like a conventional RING E3, with its N terminus recruiting Cdc53 and C-terminal RING activating Ubc12 Rub1. Dcn1's "potentiating neddylation" domain (Dcn1(P)) acts as an additional E3, reducing nonspecific Hrt1-mediated Ubc12 similar to Rub1 discharge and directing Ubc12's active site to Cdc53. Crystal structures of Dcn1(P)-Cdc53(WHB) and Ubc12 allow modeling of a catalytic complex, supported by mutational data. We propose that Dcn1's interactions with both Cdc53 and Ubc12 would restrict the otherwise flexible Hill RING-bound Ubc12 Rub1 to a catalytically competent orientation. Our data reveal mechanisms by which two E3s function synergistically to promote UBL transfer from one E2 to a target.

    Original languageEnglish
    Pages (from-to)784-796
    Number of pages13
    JournalMolecular Cell
    Volume39
    Issue number5
    DOIs
    Publication statusPublished - 10 Sep 2010

    Keywords

    • UBIQUITIN-LIGASE COMPLEX
    • ANAPHASE-PROMOTING COMPLEX
    • SACCHAROMYCES-CEREVISIAE
    • CULLIN NEDDYLATION
    • STRUCTURAL BASIS
    • ALLOSTERIC ACTIVATION
    • CONJUGATING ENZYMES
    • PROTEIN LIGASES
    • SCF FUNCTION
    • E2

    Cite this

    Scott, D. C., Monda, J. K., Grace, C. R. R., Duda, D. M., Kriwacki, R. W., Kurz, T., & Schulman, B. A. (2010). A dual E3 mechanism for Rub1 Ligation to Cdc53. Molecular Cell, 39(5), 784-796. https://doi.org/10.1016/j.molcel.2010.08.030