Cullin-RING ligase BioE3 reveals molecular-glue-induced neosubstrates and rewiring of the endogenous Cereblon ubiquitome.

ABSTRACT:

Background

The specificity of the ubiquitination process is mediated by the E3 ligases. Discriminating genuine substrates of E3s from mere interacting proteins is one of the major challenges in the field. We previously developed BioE3, a biotin-based approach that uses BirA-E3 fusions together with ubiquitin fused to a low-affinity AviTag to obtain a site-specific and proximity-dependent biotinylation of the substrates. We proved the suitability of BioE3 to identify targets of RING and HECT-type E3 ligases.

Methods

BioE3 experiments were performed in HEK293FT and U2OS stable cell lines expressing TRIPZ-bio^GEFUb transiently transfected with BirA-cereblon (CRBN). Cells were seeded using biotin-free media, followed later by a short-biotin pulse. We evaluated the appl

SUBMITTER: Merino-Cacho L 

PROVIDER: S-EPMC11841277 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature