Regulation of cullin-RING E3 ubiquitin-ligases by neddylation and dimerization

Merlet, Jorge; Burger, Julien; Gomes, José‐Eduardo; Pintard, Lionel

doi:10.1007/s00018-009-8712-7

Cited by 170 publications

(183 citation statements)

References 140 publications

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“…6C, left and middle panels). Several previous studies have also shown that CRLs take dynamic conformations (35,36), the flexibility of which may be rate-limiting for ligase activity. Dimerizations of substrate recognition proteins may influence the positioning of the substrate within the complex, as suggested by the analysis of the F-box protein Cdc4.…”

Section: Discussionmentioning

confidence: 96%

Ubiquitin Ligase Activity of Cul3-KLHL7 Protein Is Attenuated by Autosomal Dominant Retinitis Pigmentosa Causative Mutation

Kigoshi

Tsuruta

Chiba

2011

Journal of Biological Chemistry

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Substrate-specific protein degradation mediated by the ubiquitin proteasome system (UPS) is crucial for the proper function of the cell. Proteins are specifically recognized and ubiquitinated by the ubiquitin ligases (E3s) and are then degraded by the proteasome. BTB proteins act as the substrate recognition subunit that recruits their cognate substrates to the Cullin 3-based multisubunit E3s. Recently, it was reported that missense mutations in KLHL7, a BTB-Kelch protein, are related to autosomal dominant retinitis pigmentosa (adRP). However, the involvement of KLHL7 in the UPS and the outcome of the adRP causative mutations were unknown. In this study, we show that KLHL7 forms a dimer, assembles with Cul3 through its BTB and BACK domains, and exerts E3 activity. Lys-48-linked but not Lys-63-linked polyubiquitin chain co-localized with KLHL7, which increased upon proteasome inhibition suggesting that KLHL7 mediates protein degradation via UPS. An adRP-causative missense mutation in the BACK domain of KLHL7 attenuated only the Cul3 interaction but not dimerization. Nevertheless, the incorporation of the mutant as a heterodimer in the Cul3-KLHL7 complex diminished the E3 ligase activity. Together, our results suggest that KLHL7 constitutes a Cul3-based E3 and that the disease-causing mutation inhibits ligase activity in a dominant negative manner, which may lead to the inappropriate accumulation of the substrates targeted for proteasomal degradation.

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Section: Discussionmentioning

confidence: 96%

Ubiquitin Ligase Activity of Cul3-KLHL7 Protein Is Attenuated by Autosomal Dominant Retinitis Pigmentosa Causative Mutation

Kigoshi

Tsuruta

Chiba

2011

Journal of Biological Chemistry

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“…One of the recovered interactors, which encodes the full length of Nedd8 protein, attracted our attention for two reasons. First, neddylation has been well defined as an essential activation mechanism of Cullin-RING E3s 4 . The relationship between neddylation and HECT E3s has not been studied.…”

Section: Nedd8 Correlates With Smurf1 Expression In Colorectal Cancermentioning

confidence: 99%

“…In contrast, RING E3s are scaffolds that recruit E2 to the substrate to facilitate the direct transfer of ubiquitin 1 . Cullin-RING ligases (CRLs) are the most prominent class of E3s and play a pivotal role in the regulation of immunity, development, cell signalling and cell cycle 3,4 . Modification of CRLs by the ubiquitin-like Nedd8 (neural precursor cell-expressed developmentally downregulated protein 8), called neddylation, has been demonstrated to be essential for their activation 1,5 .…”

mentioning

confidence: 99%

The covalent modifier Nedd8 is critical for the activation of Smurf1 ubiquitin ligase in tumorigenesis

et al. 2014

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Neddylation, the covalent attachment of ubiquitin-like protein Nedd8, of the Cullin-RING E3 ligase family regulates their ubiquitylation activity. However, regulation of HECT ligases by neddylation has not been reported to date. Here we show that the C2-WW-HECT ligase Smurf1 is activated by neddylation. Smurf1 physically interacts with Nedd8 and Ubc12, forms a Nedd8-thioester intermediate, and then catalyses its own neddylation on multiple lysine residues. Intriguingly, this autoneddylation needs an active site at C426 in the HECT N-lobe. Neddylation of Smurf1 potently enhances ubiquitin E2 recruitment and augments the ubiquitin ligase activity of Smurf1. The regulatory role of neddylation is conserved in human Smurf1 and yeast Rsp5. Furthermore, in human colorectal cancers, the elevated expression of Smurf1, Nedd8, NAE1 and Ubc12 correlates with cancer progression and poor prognosis. These findings provide evidence that neddylation is important in HECT ubiquitin ligase activation and shed new light on the tumour-promoting role of Smurf1.

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“…Although a Ub chain is the preferred binding partner of these shuttle proteins, monoubiquitin (monoUb) has been shown to interact with them as well, albeit with lower affinity (58 -62). Moreover, as Rub1 is documented to form monomeric modifications on its targets (26,28), comparison of its binding preferences with monoUb might unlock a clue as to what distinguishes the two. To assess whether Rub1 interacts with the UBA domains of these receptors, 15 N labeled Rub1 was titrated with the UBA domains of hHR23a, human UQ1, and yeast Ddi1, and the interaction was monitored using NMR spectroscopy (see Fig.…”

Section: Rub1 Is Structurally Similar To Ubiquitin-although Rub1 Ismentioning

confidence: 99%

“…In contrast, the Rub1 pathway exclusively attaches monomeric Rub1 to substrates and performs mainly regulatory functions (26). The attachment of Rub1 (rubylation) to cullin proteins and the subsequent regulation of a multi-subunit E3 ligase, the Skp, cullin, and F-box containing complex, is probably the most prevalent, and clearly the best studied, biological function reported so far (22,23,27,28). However, some other proteins have recently been shown to be modified by Nedd8, including p53 and its E3 ligase Mdm2 (29 -31), p73 (a homolog of p53) (32), ribosomal protein L11 (33,34), epidermal growth factor receptor (EGFR) (35), caspase-7 (36,37), and parkin (38,39), though the effects are still vague, particularly as the ratio of modified to unmodified forms of each protein is extremely low.…”

mentioning

confidence: 99%

Recognition and Cleavage of Related to Ubiquitin 1 (Rub1) and Rub1-Ubiquitin Chains by Components of the Ubiquitin-Proteasome System

Singh

Zerath

Kleifeld

et al. 2012

Molecular & Cellular Proteomics

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Of all ubiquitin-like proteins, Rub1 (Nedd8 in mammals) is the closest kin of ubiquitin. We show via NMR that structurally, Rub1 and ubiquitin are fundamentally similar as well. Despite these profound similarities, the prevalence of Rub1/Nedd8 and of ubiquitin as modifiers of the proteome is starkly different, and their attachments to specific substrates perform different functions. Recently, some proteins, including p53, p73, EGFR, caspase-7, and Parkin, have been shown to be modified by both Rub1/ Nedd8 and ubiquitin within cells. To understand whether and how it might be possible to distinguish among the same target protein modified by Rub1 or ubiquitin or both, we examined whether ubiquitin receptors can differentiate between Rub1 and ubiquitin. Surprisingly, Rub1 interacts with proteasome ubiquitin-shuttle proteins comparably to ubiquitin but binds more weakly to a proteasomal ubiquitin receptor Rpn10. We identified Rub1-ubiquitin heteromers in yeast and Nedd8-Ub heteromers in human cells. We validate that in human cells and in vitro, human Rub1 (Nedd8) forms chains with ubiquitin where it acts as a chain terminator. Interestingly, enzymatically assembled K48-linked Rub1-ubiquitin heterodimers are recognized by various proteasomal ubiquitin shuttles and receptors comparably to K48-linked ubiquitin homodimers. Furthermore, these heterologous chains are cleaved by COP9 signalosome or 26S proteasome. A derubylation function of the proteasome expands the repertoire of its enzymatic activities. In contrast, Rub1 conjugates may be somewhat resilient to the actions of other canonical deubiquitinating enzymes. Taken together, these findings suggest that once Rub1/Nedd8 is channeled into ubiquitin pathways, it is recognized essentially like The selective degradation of many proteins in eukaryotic cells is a highly specific and irreversible process required to perform vital cellular functions such as cell cycle progression (1, 2), differentiation and development (3, 4), and transcriptional control (5). One of the major pathways involved in this selective degradation is the ubiquitin-proteasome pathway. In this pathway, ubiquitin (Ub), a 76-amino-acid protein, is attached to the substrate, and this is followed by the recognition and degradation of the substrate by a protein complex collectively known as proteasome (6 -8).The attachment of Ub (ubiquitination) to a substrate protein is achieved via a cascade of enzymatic reactions (involving E1, E2, and E3 enzymes) that results in the formation of an isopeptide bond between the C-terminal glycine of Ub and a lysine residue of the substrate (9 -11). Sequential repetition of this cascade can result in the attachment of a chain of Ub molecules (polyubiquitin) to the substrate protein. The length and topology of the polyubiquitin (polyUb) tag decide the fate of the substrate protein. For example, we and others have shown that a K48-linked tetraubiquitin (tetraUb) chain that acts as a proteasomal degradation signal forms a "closed" structure (12, 13), whereas a K63-linked Ub...

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Regulation of cullin-RING E3 ubiquitin-ligases by neddylation and dimerization

Cited by 170 publications

References 140 publications

Ubiquitin Ligase Activity of Cul3-KLHL7 Protein Is Attenuated by Autosomal Dominant Retinitis Pigmentosa Causative Mutation

Ubiquitin Ligase Activity of Cul3-KLHL7 Protein Is Attenuated by Autosomal Dominant Retinitis Pigmentosa Causative Mutation

The covalent modifier Nedd8 is critical for the activation of Smurf1 ubiquitin ligase in tumorigenesis

Recognition and Cleavage of Related to Ubiquitin 1 (Rub1) and Rub1-Ubiquitin Chains by Components of the Ubiquitin-Proteasome System

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