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

Jingwei Zeng; Ana Filipa Santos; Aamir S. Mukadam; Mariana Osswald; David A. Jacques; Claire F. Dickson; Stephen H. McLaughlin; Christopher M. Johnson; L. Kiss; Jakub Lupták; Nadine Renner; Marina Vaysburd; William A. McEwan; Eurico Morais‐de‐Sá; Dean Clift; Leo C. James

doi:10.1038/s41594-021-00560-2

Molecular BiologyNature Structural & Molecular BiologyFeb 25, 2021

Peer-reviewed journal article

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

Jingwei Zeng iD, Ana Filipa Santos iD, Aamir S. Mukadam iD, Mariana Osswald iD, David A. Jacques iD, Claire F. Dickson iD, Stephen H. McLaughlin iD, Christopher M. Johnson iD, L. Kiss iD, Jakub Lupták iD, Nadine Renner, Marina Vaysburd, William A. McEwan iD, Eurico Morais‐de‐Sá iD, Dean Clift iD, Leo C. James iD

Abstract

Trim-Away is a powerful new technology that exploits off-the-shelf antibodies and the E3 RING ligase and cytosolic antibody receptor TRIM21 to carry out rapid protein depletion. How TRIM21 is catalytically-activated upon substrate engagement during either its normal immune function or when re-purposed for targeted protein degradation is unknown. Here we show that a mechanism of substrate-induced clustering triggers intermolecular dimerization of the RING domain to switch on the ubiquitination activity of TRIM21 and induce an antiviral response or drive Trim-Away. We harness this mechanism to expand the Trim-Away toolbox with highly-active TRIM21-nanobody chimeras that can also be controlled optogenetically. This work provides a mechanism for cellular activation of TRIM RING ligases and has important implications for targeted protein degradation technologies.

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