Allosteric Activation of E2-RING Finger-Mediated Ubiquitylation by a Structurally Defined Specific E2-Binding Region of gp78

Das, Ranabir; Mariano, Jennifer; Tsai, Ya Chea; Kalathur, Ravi C.; Kostova, Zlatka; Li, Jess; Tarasov, Sergey G.; McFeeters, Robert L.; Altieri, Amanda S.; Ji, Xinhua; Byrd, R. Andrew; Weissman, Allan M.

doi:10.1016/j.molcel.2009.05.010

Cited by 148 publications

(280 citation statements)

References 40 publications

(64 reference statements)

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“…1C). This surface, comprising residues on the N-terminal helix (H1), loop 4, and loop 7 (L4 and L7), is the canonical E3 interaction surface through which UbcH7 interacts with both HECT and RING E3 ligases (11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23). A highly conserved phenylalanine residue in L4 (F63 in UbcH7) seems to be the determinant for HECT E3 recognition.…”

Section: Resultsmentioning

confidence: 99%

Crystal structures of two bacterial HECT-like E3 ligases in complex with a human E2 reveal atomic details of pathogen-host interactions

Lin¹,

Diao

Chen

2012

Proc. Natl. Acad. Sci. U.S.A.

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In eukaryotes, ubiquitination is an important posttranslational process achieved through a cascade of ubiquitin-activating (E1), conjugating (E2), and ligase (E3) enzymes. Many pathogenic bacteria deliver virulence factors into the host cell that function as E3 ligases. How these bacterial "Trojan horses" integrate into the eukaryotic ubiquitin system has remained a mystery. Here we report crystal structures of two bacterial E3s, Salmonella SopA and Escherichia coli NleL, both in complex with human E2 UbcH7. These structures represent two distinct conformational states of the bacterial E3s, supporting the necessary structural rearrangements associated with ubiquitin transfer. The E2-interacting surface of SopA and NleL has little similarity to those of eukaryotic E3s. However, both bacterial E3s bind to the canonical surface of E2 that normally interacts with eukaryotic E3s. Furthermore, we show that a glutamate residue on E3 is involved in catalyzing ubiquitin transfer from E3 to the substrate, but not from E2 to E3. Together, these results provide mechanistic insights into the ubiquitin pathway and a framework for understanding molecular mimicry in bacterial pathogenesis.crystallography | microbe-host interaction | NleL | SopA | ubiquitination I n order to coexist with eukaryotic hosts, bacterial pathogens have evolved mechanisms to alter the physiological and immune response of the host. For example, many gram-negative bacteria use the type III or type IV secretion system to deliver a large number of virulence factors into the host cell cytosol. These virulence factors, also known as effector proteins, play vital roles in bacterial attachment, entry, and survival. They modulate numerous host cellular functions such as cytoskeleton dynamics, gene expression, and posttranslational modification. Understanding how bacterial virulence factors exert their effects on host cell processes will offer new insights into eukaryotic host cell physiology and possibly lead to new approaches to antimicrobial therapy.The ubiquitin (Ub) pathway is one of the host systems that is hijacked by bacterial pathogens (1, 2). In eukaryotes ubiquitination is central to many processes such as cell cycle, immune response, and DNA damage tolerance (3-6). The covalent attachment of Ub to a target protein requires the sequential actions of Ub-activating enzymes (E1), conjugating enzymes (E2), and ligases (E3). An Ub molecule is first attached to E1 through a thioester bond upon ATP hydrolysis and is subsequently transferred to the active site cysteine residue in E2. The E3 ligase is often needed to transfer Ub from E2 to a protein substrate. There are two major types of E3 ligases: the RING E3s that function as scaffold to bring E2 and substrate into proximity and HECT E3s that form a thioester intermediate with the Ub before transferring it to the substrate. Ubiquitination is absent in prokaryotes. However, some pathogenic bacteria deliver E3 ligases that interfere with the eukaryotic Ub pathway. These bacterial-encoded E3 ligases have litt...

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Section: Resultsmentioning

confidence: 99%

Crystal structures of two bacterial HECT-like E3 ligases in complex with a human E2 reveal atomic details of pathogen-host interactions

Lin¹,

Diao

Chen

2012

Proc. Natl. Acad. Sci. U.S.A.

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“…The low RING-RBL/UBE2E1 affinity is consistent with that in other RING/E2 interactions (36, 44 -46). However, it has recently been shown that allosteric activation of the E2 by regions of the E3 further apart in sequence from the RING domain can decrease the E2/E3 dissociation constant substantially (46), which may also be the case for Ro52 and other TRIM proteins.…”

Section: Discussionmentioning

confidence: 99%

Anti-Ro52 Autoantibodies from Patients with Sjögren's Syndrome Inhibit the Ro52 E3 Ligase Activity by Blocking the E3/E2 Interface

Espinosa

Hennig

Ambrosi

et al. 2011

Journal of Biological Chemistry

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Ro52 (TRIM21)is an E3 ligase of the tripartite motif family that negatively regulates proinflammatory cytokine production by ubiquitinating transcription factors of the interferon regulatory factor family. Autoantibodies to Ro52 are present in patients with lupus and Sjögren's syndrome, but it is not known if these autoantibodies affect the function of Ro52. To address this question, the requirements for Ro52 E3 ligase activity were first analyzed in detail. Scanning a panel of E2 ubiquitin-conjugating enzymes, we found that UBE2D1-4 and UBE2E1-2 supported the E3 ligase activity of Ro52 and that the E3 ligase activity of Ro52 was dependent on its RING domain. We also found that the N-terminal extensions in the class III E2 enzymes affected their interaction with Ro52. Although the N-terminal extension in UBE2E3 made this E2 enzyme unable to function together with Ro52, the N-terminal extensions in UBE2E1 and UBE2E2 allowed for a functional interaction with Ro52. AntiRo52-positive patient sera and affinity-purified anti-RING domain autoantibodies inhibited the E3 activity of Ro52 in ubiquitination assays. Using NMR, limited proteolysis, ELISA, and Ro52 mutants, we mapped the interactions between Ro52, UBE2E1, and anti-Ro52 autoantibodies. We found that antiRo52 autoantibodies inhibited the E3 ligase activity of Ro52 by sterically blocking the E2/E3 interaction between Ro52 and UBE2E1. Our data suggest that anti-Ro52 autoantibodies binding the RING domain of Ro52 may be actively involved in the pathogenesis of rheumatic autoimmune disease by inhibiting Ro52-mediated ubiquitination.Ro52 (TRIM21) is an interferon-inducible protein frequently targeted by autoantibodies in patients with primary Sjögren's syndrome (SS) 4 and systemic lupus erythematosus (SLE) (1, 2). As a member of the tripartite motif (TRIM) protein family (3), Ro52 is characterized by a RING domain (4), a B-box type 2 motif (5), and a coiled-coil region. A B30.2 domain is located in the C-terminal region (6). Several TRIM family members, including Ro52, have been identified as E3 ubiquitin ligases involved in the ubiquitination process (7-10). Ubiquitination is a covalent post-translational modification of proteins by the 76-amino acid residue polypeptide ubiquitin, leading either to proteolysis in the proteasome, internalization from membranes, or functional alteration (11,12). After activation by a ubiquitin-activating enzyme (E1), ubiquitin is transferred to a ubiquitin-conjugating enzyme (E2) followed by a ubiquitin ligase (E3)-mediated transfer of ubiquitin from the E2 to a target protein (13). Ro52 mediates ubiquitination of several members of the interferon regulatory factor (IRF) transcription factor family, thereby regulating cytokine production (14 -17). Gene disruption of Ro52 leads to increased production of proinflammatory cytokines by embryonic fibroblasts and immune cells after Toll-like receptor activation, suggesting that Ro52 negatively regulates IRF activity (16,17).Autoantibodies to Ro52 can appear years before tissue damage is ...

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“…Subsequently, AMFR catalyzes K27-linked polyubiquitylation of STING, which serves as a scaffold for the recruitment and activation of TBK1 (Wang et al, 2014). Of note, it has been shown that AMFR interacts with the E2 enzyme UBE2G2 through a specialized binding region on AMFR, and that this interaction is a prerequisite for processive assembly of K48-linked ubiquitin chains on ER-associated degradation (ERAD) substrates (Das et al, 2013(Das et al, , 2009). It would be exciting to uncover the molecular mechanisms of how AMFR can also assemble K27-linked ubiquitin chains.…”

Section: K11 Linkages In Heterotypic Ubiquitin Conjugates -A Powerfulmentioning

confidence: 99%

Ubiquitin chain diversity at a glance

Akutsu

Đikić

Bremm

2016

Journal of Cell Science

380

367

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Ubiquitin plays an essential role in modulating protein functions, and deregulation of the ubiquitin system leads to the development of multiple human diseases. Owing to its molecular features, ubiquitin can form various homo-and heterotypic polymers on substrate proteins, thereby provoking distinct cellular responses. The concept of multifaceted ubiquitin chains encoding different functions has been substantiated in recent years. It has been established that all possible ubiquitin linkage types are utilized for chain assembly and propagation of specific signals in vivo. In addition, branched ubiquitin chains and phosphorylated ubiquitin molecules have been put under the spotlight recently. The development of novel technologies has provided detailed insights into the structure and function of previously poorly understood ubiquitin signals. In this Cell Science at a Glance article and accompanying poster, we provide an update on the complexity of ubiquitin chains and their physiological relevance.

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Allosteric Activation of E2-RING Finger-Mediated Ubiquitylation by a Structurally Defined Specific E2-Binding Region of gp78

Cited by 148 publications

References 40 publications

Crystal structures of two bacterial HECT-like E3 ligases in complex with a human E2 reveal atomic details of pathogen-host interactions

Crystal structures of two bacterial HECT-like E3 ligases in complex with a human E2 reveal atomic details of pathogen-host interactions

Anti-Ro52 Autoantibodies from Patients with Sjögren's Syndrome Inhibit the Ro52 E3 Ligase Activity by Blocking the E3/E2 Interface

Ubiquitin chain diversity at a glance

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