The molecular mechanisms that drive intracellular neutralization by the antibody-receptor and RING E3 ligase TRIM21

Kiss, Leo; James, Leo C.

doi:10.1016/j.semcdb.2021.11.005

Cited by 13 publications

(15 citation statements)

References 108 publications

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“…This and subsequent work revealed that TRIM5α-mediated RT inhibition and innate signaling depend on two specialized E2 proteins, UBE2W and UBE2N, and implicated the N-terminal autoubiquitylation of TRIM5α with K63-linked polyubiquitin chains in its antiviral activity 36 – 38 . Antibody-dependent intracellular neutralization of viruses by another TRIM family member, TRIM21, was shown to similarly require UBE2W- and UBE2N-mediated autoubiquitylation 39 – 41 . Dependence of TRIM5α and TRIM21 on two distinct E2s for priming and extension of the polyubiquitin chain bears resemblance to E3 mechanisms previously described for other, non-TRIM, ubiquitin ligases 42 – 45 .…”

Section: Introductionmentioning

confidence: 99%

Structural and functional asymmetry of RING trimerization controls priming and extension events in TRIM5α autoubiquitylation

Herkules

Taylor

et al. 2022

Nat Commun

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TRIM5α is an E3 ubiquitin ligase of the TRIM family that binds to the capsids of primate immunodeficiency viruses and blocks viral replication after cell entry. Here we investigate how synthesis of K63-linked polyubiquitin is upregulated by transient proximity of three RING domains in honeycomb-like assemblies formed by TRIM5α on the surface of the retroviral capsid. Proximity of three RINGs creates an asymmetric arrangement, in which two RINGs form a catalytic dimer that activates E2-ubiquitin conjugates and the disordered N-terminus of the third RING acts as the substrate for N-terminal autoubiquitylation. RING dimerization is required for activation of the E2s that contribute to the antiviral function of TRIM5α, UBE2W and heterodimeric UBE2N/V2, whereas the proximity of the third RING enhances the rate of each of the two distinct steps in the autoubiquitylation process: the initial N-terminal monoubiquitylation (priming) of TRIM5α by UBE2W and the subsequent extension of the K63-linked polyubiquitin chain by UBE2N/V2. The mechanism we describe explains how recognition of infection-associated epitope patterns by TRIM proteins initiates polyubiquitin-mediated downstream events in innate immunity.

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Section: Introductionmentioning

confidence: 99%

Structural and functional asymmetry of RING trimerization controls priming and extension events in TRIM5α autoubiquitylation

Herkules

Taylor

et al. 2022

Nat Commun

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“…Importantly, while autoubiquitination explains why TRIM ligases are degraded upon activation, it does not provide a direct mechanism for substrate degradation. Whether substrates are also modified with ubiquitin during their cellular degradation is unknown, despite attempts to detect this 2,16 , and so because of the absence of such data it has been proposed that ligase autoubiquitination alone may drive proteasome recruitment, resulting in degradation of the entire TRIM:substrate complex 2,25,43,44 .…”

Section: Introductionmentioning

confidence: 99%

Trim-Away degrades lysine-less substrates without requiring ligase autoubiquitination

Kiss

Rhinesmith

Dickson

et al. 2022

Preprint

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TRIM proteins are the largest family of E3 ligases in mammals. They include the intracellular antibody receptor TRIM21, which is responsible for mediating targeted protein degradation during Trim-Away. Despite their importance, the ubiquitination mechanism of TRIM ligases has remained elusive. Here we show that while TRIM21 activation results in ubiquitination of both ligase and substrate, autoubiquitination is regulatory and not required for substrate degradation. Substrate binding stimulates N-terminal RING autoubiquitination by the E2 Ube2W, but when inhibited by N-terminal acetylation this prevents neither substrate ubiquitination nor degradation and has no impact on TRIM21 antiviral activity. Instead, uncoupling ligase and substrate degradation prevents ligase recycling and extends functional persistence in cells. Substrate ubiquitination mediates degradation but Trim-Away efficiently degrades lysine-less substrates, suggesting a non-canonical ubiquitination mechanism explains its broad substrate specificity

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“…Tripartite motif containing protein 21 (TRIM21) is an intracellular immunoglobulin receptor and E3 ubiquitin ligase that functions in the innate immune response (Kiss and James, 2021). When non-enveloped viruses enter a cell, they bring with them capsid-bound serum antibodies (Mallery et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

“…The domain architecture and enzymology of TRIM21 suggest that activation of its E3 ubiquitin ligase activity requires higher order assembly (Kiss and James, 2021). TRIM21 contains an N-terminal RING domain and an internal coiled-coil domain that mediates anti-parallel TRIM21 homodimerization, placing the RING domains at opposite ends of a TRIM21 dimer (Figure 1A).…”

Section: Introductionmentioning

confidence: 99%

“…The K63-linked polyubiquitin chain on TRIM21 has been proposed to trigger immune signaling as well as the proteasomal degradation not only of TRIM21, but also of antibody and substrate, whose ubiquitination has not been detected and is assumed not to occur (Figure 1B) (Fletcher et al, 2015). The destruction of proteins without direct ubiquitination is unconventional, but would allow TRIM21 to destroy a potentially unlimited number of substrates, as long as they are bound by a critical number of antibodies (Kiss and James, 2021; Zeng et al, 2021). Because the proteasome generally does not act on K63-linked polyubiquitin chains, it has been proposed that this modification scaffolds subsequent formation of K48 chains, the canonical proteasomal degradation signal.…”

Section: Introductionmentioning

confidence: 99%

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Cell-free Trim-Away reveals the mechanism of antibody-mediated protein degradation by TRIM21

Mevissen

Prasad

Walter

2022

Preprint

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TRIM21 is a cytosolic antibody receptor and E3 ubiquitin ligase that promotes destruction of a broad range of pathogens. TRIM21 also underlies the antibody-dependent protein targeting method Trim-Away. Current evidence suggests that TRIM21 binding to antibodies leads to formation of a self-anchored K63 ubiquitin chain on the N-terminus of TRIM21 that triggers the destruction of TRIM21, antibody, and target protein. Here, we report that addition of antibody and TRIM21 to Xenopus egg extracts promotes efficient degradation of endogenous target proteins, establishing cell-free Trim-Away as a powerful tool to interrogate protein function. Chemical methylation of TRIM21 had no effect on target proteolysis, whereas deletion of all lysine residues in targets abolished their ubiquitination and proteasomal degradation. These results demonstrate that target protein but not TRIM21 polyubiquitination is required for Trim-Away, and they suggest that current models of TRIM21 function should be fundamentally revised.

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The molecular mechanisms that drive intracellular neutralization by the antibody-receptor and RING E3 ligase TRIM21

Cited by 13 publications

References 108 publications

Structural and functional asymmetry of RING trimerization controls priming and extension events in TRIM5α autoubiquitylation

Structural and functional asymmetry of RING trimerization controls priming and extension events in TRIM5α autoubiquitylation

Trim-Away degrades lysine-less substrates without requiring ligase autoubiquitination

Cell-free Trim-Away reveals the mechanism of antibody-mediated protein degradation by TRIM21

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