Cullin neddylation and substrate-adaptors counteract SCF inhibition by the CAND1-like protein Lag2 in Saccharomyces cerevisiae

Edyta Siergiejuk, Daniel C. Scott, Brenda A. Schulman, Kay Hofmann, Thimo Kurz, Matthias Peter

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    26 Citations (Scopus)

    Abstract

    Cullin-based E3 ubiquitin ligases are activated through covalent modification of the cullin subunit by the ubiquitin-like protein Nedd8. Cullin neddylation dissociates the ligase assembly inhibitor Cand1, and promotes E2 recruitment and ubiquitin transfer by inducing a conformational change. Here, we have identified and characterized Lag2 as a likely Saccharomyces cerevisiae orthologue of mammalian Cand1. Similar to Cand1, Lag2 directly interacts with non-neddylated yeast cullin Cdc53 and prevents its neddylation in vivo and in vitro. Binding occurs through a conserved C-terminal beta-hairpin structure that inserts into the Skp1-binding pocket on the cullin, and an N-terminal motif that covers the neddylation lysine. Interestingly, Lag2 is itself neddylated in vivo on a lysine adjacent to this N-terminal-binding site. Overexpression of Lag2 inhibits Cdc53 activity in strains defective for Skp1 or neddylation functions, implying that these activities are important to counteract Lag2 in vivo. Our results favour a model in which binding of substrate-specific adaptors triggers release of Cand1/Lag2, whereas subsequent neddylation of the cullin facilitates the removal and prevents re-association of Lag2/Cand1. The EMBO Journal (2009) 28, 3845-3856. doi: 10.1038/emboj.2009.354; Published online 26 November 2009

    Original languageEnglish
    Pages (from-to)3845-3856
    Number of pages12
    JournalEMBO Journal
    Volume28
    Issue number24
    DOIs
    Publication statusPublished - 16 Dec 2009

    Keywords

    • Cand1
    • cell cycle
    • cullin
    • Nedd8
    • ubiquitin
    • UBIQUITIN LIGASE
    • COP9 SIGNALOSOME
    • E3 LIGASE
    • COMPLEX REVEALS
    • F-BOX
    • NEDD8
    • CONJUGATION
    • ACTIVATION
    • MECHANISMS
    • PATHWAY

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