Substrate-induced clustering activates Trim-Away of pathogens and proteins

Zeng, Jingwei; Santos, Ana Filipa; Mukadam, Aamir S.; Osswald, Mariana; Lupták, Jakub; Jacques, D.A.; Dickson, Claire F.; Renner, Nadine; Johnson, C.M.; Vaysburd, Marina; McEwan, William A.; Morais-de-Sá, Eurico; Clift, Dean; James, Leo C.

doi:10.1101/2020.07.28.225359

Cited by 2 publications

(2 citation statements)

References 82 publications

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“…TRIM21 forms inactive dimers in which a central antiparallel coiled coil positions the two RING domains at the two ends of the dimer. Upon binding to multimeric substrates, RING domains from neighboring TRIM21 dimerizes, leading to the activation of its enzymatic activity 37 . In line with substrate-induced activation of TRIM21, we present two classes of TRIM21-based degraders with a high degree of selectivity towards multimeric neo-substrates.…”

Section: Discussionmentioning

confidence: 99%

“…TRIM21 has been exploited in the Trim-Away technology, which uses antibodies to direct TRIM21 to degrade intracellular proteins 36 . A recent mechanistic study revealed that substrateinduced clustering triggers intermolecular dimerization of the RING domains of TRIM21 to switch on its enzymatic activity 37 . Interestingly, NUP98 APD , the degron recognized by TRIM21, is also embedded in the multimeric nuclear pore complex (Figure 5K).…”

Section: Trim21-based Protacs Selectively Degrade Multimeric Proteins...mentioning

confidence: 99%

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Selective degradation of multimeric proteins via chemically induced proximity to TRIM21

Lu,

Cheng,

Xue

et al. 2024

Preprint

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Targeted protein degradation (TPD) has emerged as an effective strategy to eliminate disease-causing proteins by inducing their interactions with the protein degradation machinery. First-generation TPD agents exploit a limited set of broadly expressed E3 ubiquitin ligases with constitutive activity, forbidding their application to proteins requiring higher levels of targeting selectivity. Here, by phenotype-based screening, we discovered that the antipsychotic drug acepromazine possesses interferon-enhanced cytotoxicity towards cancer cell lines expressing high levels of aldo-keto reductases 1C. These enzymes convert acepromazine into its stereo-selective metabolite (S)-hydroxyl-acepromazine, which recruits the interferon-induced E3 ubiquitin ligase TRIM21 to the vicinity of the nuclear pore complex, resulting in the degradation of nuclear pore proteins. Co-crystal structures of acepromazine and derivatives in complex with the PRYSPRY domain of TRIM21 revealed a ligandable pocket, which was exploited for designing heterobifunctional degraders. The resulting chemicals selectively degrade multimeric proteins— such as those in biomolecular condensates—without affecting monomeric proteins, consistent with the requirement of substrate-induced clustering for TRIM21 activation. As aberrant protein assemblies have been causally linked to diseases such as neurodegeneration, autoimmunity, and cancer, our findings highlight the potential of TRIM21-based multimer-selective degraders as a strategy to tackle the direct causes of these diseases.

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Section: Discussionmentioning

confidence: 99%

Section: Trim21-based Protacs Selectively Degrade Multimeric Proteins...mentioning

confidence: 99%

Selective degradation of multimeric proteins via chemically induced proximity to TRIM21

Lu,

Cheng,

Xue

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

RING domains act as both substrate and enzyme in a catalytic arrangement to drive self-anchored ubiquitination

et al. 2021

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Attachment of ubiquitin (Ub) to proteins is one of the most abundant and versatile of all posttranslational modifications and affects outcomes in essentially all physiological processes. RING E3 ligases target E2 Ub-conjugating enzymes to the substrate, resulting in its ubiquitination. However, the mechanism by which a ubiquitin chain is formed on the substrate remains elusive. Here we demonstrate how substrate binding can induce a specific RING topology that enables self-ubiquitination. By analyzing a catalytically trapped structure showing the initiation of TRIM21 RING-anchored ubiquitin chain elongation, and in combination with a kinetic study, we illuminate the chemical mechanism of ubiquitin conjugation. Moreover, biochemical and cellular experiments show that the topology found in the structure can be induced by substrate binding. Our results provide insights into ubiquitin chain formation on a structural, biochemical and cellular level with broad implications for targeted protein degradation.

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Substrate-induced clustering activates Trim-Away of pathogens and proteins

Cited by 2 publications

References 82 publications

Selective degradation of multimeric proteins via chemically induced proximity to TRIM21

Selective degradation of multimeric proteins via chemically induced proximity to TRIM21

RING domains act as both substrate and enzyme in a catalytic arrangement to drive self-anchored ubiquitination

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