Mechanistic Insights Into the Assembly of Functional CRL3 Dimeric Complexes.

Abstract

The assembly of Cullin3-based RING E3 ubiquitin ligase (CRL3) complexes is orchestrated in two consecutive steps: the formation of the dimeric BTB domain core and the recruitment of CUL3-RBX1 subunits. Each step is tightly regulated to ensure the formation of complete and functional dimeric CRL3s. The first assembly step is regulated by two mechanisms: "co-co assembly" and proteasome-dependent degradation of aberrant heterodimers. The second step is facilitated by a conserved CUL3 N-terminal assembly (NA) motif. The CUL3 NA motif contributes to the assembly of CRL3s in two aspects: interacting with both BTB domain-containing protein protomers to facilitate complete dimeric assembly, and enhancing the stability of CRL3s by overcoming the tensions generated by conformational entropy during ubiquitin transfer. Given that all Cullin proteins contain N-terminal extensions, we postulate that these extensions, similar to the CUL3 NA motif-contributed assembly, play an important role in the functional regulation of CRLs and thus warrant further investigation.