60th residues of ubiquitin and Nedd8 are located out of E2‐binding surfaces, but are important for K48 ubiquitin‐linkage

Choi, Yun‐Seok; Jeon, Young Ho; Ryu, Kyoung‐Seok; Cheong, Chaejoon

doi:10.1016/j.febslet.2009.09.034

Cited by 21 publications

(15 citation statements)

References 30 publications

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“…These results suggest that Ube2g1, Ube2r1, and Ube2k, which are known to generate Lys-48-linked polyubiquitin efficiently without E3, seem to use a different mechanism to conjugate acceptor and donor ubiquitin. Other evidence supporting this idea is that the UB N60A mutant displayed different Lys-48-ubiquitylation activities as an acceptor for Ube2k or Ube2g1, showing a severe defect with Ube2k, whereas ubiquitylation by Ube2g1 was relatively unaffected by the ubiquitin N60A mutation (12).…”

Section: Discussionmentioning

confidence: 72%

“…Interestingly, CSP signals from the acidic loop region of Ube2g1 (Loop) were clearly identified when ubiquitin was added (Fig. 4, A (12). The acidic loop in Ube2g1 is similar to the previously characterized weak ubiquitin-binding site 1 (UBS1) in the C-terminal tail of Ube2r1 (DLFYDDYYED) (26), which also consists of hydrophobic and acidic residues (Fig.…”

Section: Two Aromatic Residues (Tyr-102 and Tyr-104) In The Ube2g1mentioning

confidence: 74%

“…Work from our group previously showed that, unlike full-length Ube2r1, the Ube2r1 core (residues 1-183; Ube2r1C) has a decreased capability to synthesize Lys-48 polyubiquitin chains efficiently in the absence of an E3 (8). Ube2d2, which does not have an acidic loop, also is unable to synthesize Lys-48 polyubiquitin in the absence of an E3 ligase (8) despite having ubiquitin binding affinity almost identical to that of Ube2g1 as assessed by NMR CSP experiments using [ 15 N]UB (12). In this study we focused on the acidic loops and ubiquitin binding activities of Ube2g1 and Ube2r1C.…”

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confidence: 99%

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Differential Ubiquitin Binding by the Acidic Loops of Ube2g1 and Ube2r1 Enzymes Distinguishes Their Lys-48-ubiquitylation Activities

Choi

Lee

et al. 2015

Journal of Biological Chemistry

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Background:The acidic loops of E2 enzymes are important for their Lys-48-ubiquitylation activity. Results: The presence of Tyr residues modulates the ubiquitin binding property of the acidic loop. Conclusion: A proper interaction of the acidic loop with the attached donor ubiquitin is important for Lys-48-ubiquitylation activity. Significance: One of molecular bases to study the mechanism of Lys-48-ubiquitylation is provided.

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Section: Discussionmentioning

confidence: 72%

Section: Two Aromatic Residues (Tyr-102 and Tyr-104) In The Ube2g1mentioning

confidence: 74%

mentioning

confidence: 99%

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Differential Ubiquitin Binding by the Acidic Loops of Ube2g1 and Ube2r1 Enzymes Distinguishes Their Lys-48-ubiquitylation Activities

Choi

Lee

et al. 2015

Journal of Biological Chemistry

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“…The overall structure of NEDD8 can be subdivided into a C-terminal tail and a globular ubiquitin-fold domain (Figure 1B). The tail is flexible in solution 26 , adopts different extended structures upon interaction with neddylation and deneddylation enzymes, and, similarly to ubiquitin, ends with a Gly-Gly sequence that becomes covalently bonded to targets 27–29 . The globular domain is characterized by four β-sheets interspersed by one α- and two 3 10 -helices (helix 1, 2, and 3 respectively).…”

Section: Structure Of the Ubiquitin-like Protein Nedd8mentioning

confidence: 99%

Protein neddylation: beyond cullin–RING ligases

Enchev

Schulman

Peter

2014

Nat Rev Mol Cell Biol

486

513

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NEDD8 is a ubiquitin-like protein that activates the largest ubiquitin E3 ligase family, the cullin RING ligases. Many non-cullin neddylation targets have been proposed in recent years. However, overexpression of exogenous NEDD8 can trigger NEDD8 conjugation through the ubiquitylation machinery, which makes validating potential NEDD8 targets challenging. Here we re-evaluate these studies in light of the current understanding of the neddylation pathway, and suggest criteria for the identification of genuine neddylation substrates under homeostatic conditions. We describe the biological processes that might be regulated by non-cullin neddylation, and the utility of neddylation inhibitors for research and as potential therapies. Understanding the biological significance of non-cullin neddylation is an exciting research prospect primed to reveal fundamental insights.

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“…2 ), and the equilibrium binding constants were obtained by fitting the CSP data to the simple binding equation (24).…”

Section: Methodsmentioning

confidence: 99%

The Human Cdc34 Carboxyl Terminus Contains a Non-covalent Ubiquitin Binding Activity That Contributes to SCF-dependent Ubiquitination

Choi

Jeong

et al. 2010

Journal of Biological Chemistry

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Cdc34 is an E2 ubiquitin-conjugating enzyme that functions in conjunction with SCF (Skp1⅐Cullin 1⅐F-box) E3 ubiquitin ligase to catalyze covalent attachment of polyubiquitin chains to a target protein.Here we identified direct interactions between the human Cdc34 C terminus and ubiquitin using NMR chemical shift perturbation assays. The ubiquitin binding activity was mapped to two separate Cdc34 C-terminal motifs (UBS1 and UBS2) that comprise residues 206-215 and 216-225, respectively. UBS1 and UBS2 bind to ubiquitin in the proximity of ubiquitin Lys 48 and C-terminal tail, both of which are key sites for conjugation. When bound to ubiquitin in one orientation, the Cdc34 UBS1 aromatic residues (Phe Ubiquitination is a complex protein modification reaction that produces a covalent isopeptide bond linking the ubiquitin Gly 76 carboxyl end to the ⑀-amino group of a lysine residue within a substrate protein (1). One of the hallmarks of this modification is that substrates are frequently attached with polyubiquitin chains, formed by isopeptide linkage joining the ubiquitin Gly 76 carboxyl end and one of its seven internal lysine residues. Three enzymatic activities typically participate in the ubiquitination reaction, including a ubiquitin-activating enzyme (E1) that forms an E1-Sϳubiquitin thiol ester complex in an ATP hydrolysis-dependent manner, a ubiquitin-conjugating enzyme (E2) capable of tethering a ubiquitin in thiol ester form that is transferred from E1, and a ubiquitin-ligase (E3). RING finger domain-containing E3s orchestrate the transfer of ubiquitin to a substrate by recruiting both the target protein and E2-Sϳubiquitin thiol ester and by helping position the attacking substrate lysine and the E2-bound donor ubiquitin optimally for conjugation (2). SCF (Skp1⅐Cullin⅐F-box) is a four-subunit RING E3 ligase complex with CUL1 (cullin 1) as scaffold that at its N terminus tethers the Skp1⅐F-box protein complex to target a substrate and utilizes its C terminus to dock the ROC1/Rbx1/Hrt1 RING finger subunit for recruiting an E2 ubiquitin-conjugating enzyme (3-9). SCF-mediated polyubiquitination is activated by conjugation of Nedd8, a ubiquitin-like protein, to human CUL1 at lysine 720 (10, 11), most likely due to Nedd8-imposed conformational changes in CUL1 (12), thereby relieving autoinhibitory interactions between the extreme C terminus of CUL1 and ROC1 (13).Accumulating evidence demonstrates a role for the Cdc34 E2 in SCF-dependent ubiquitination. Yeast Cdc34 (Ubc3) is essential for viability and is required for SCF Cdc4 -dependent degradation of the cyclin-dependent kinase inhibitor Sic1 (see Refs. 3 and 4 and references therein). This regulation has been recapitulated by in vitro experiments that reconstitute the SCF Cdcc4 -dependent and Cdc34-catalyzed ubiquitination of Sic1 and the degradation of the modified substrate by the 26 S proteasome (3,4,14). Human Cdc34 (hCdc34) 5 is highly conserved as it is

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60th residues of ubiquitin and Nedd8 are located out of E2‐binding surfaces, but are important for K48 ubiquitin‐linkage

Cited by 21 publications

References 30 publications

Differential Ubiquitin Binding by the Acidic Loops of Ube2g1 and Ube2r1 Enzymes Distinguishes Their Lys-48-ubiquitylation Activities

Differential Ubiquitin Binding by the Acidic Loops of Ube2g1 and Ube2r1 Enzymes Distinguishes Their Lys-48-ubiquitylation Activities

Protein neddylation: beyond cullin–RING ligases

The Human Cdc34 Carboxyl Terminus Contains a Non-covalent Ubiquitin Binding Activity That Contributes to SCF-dependent Ubiquitination

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