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Cryo-EM structures of Gid12-bound GID E3 reveal steric blockade as a mechanism inhibiting substrate ubiquitylation

ABSTRACT: Protein degradation, a major eukaryotic response to cellular signals, is subject to numerous layers of regulation. In yeast, the evolutionarily conserved GID E3 ligase mediates glucose-induced degradation of fructose-1,6-bisphosphatase (Fbp1) and other gluconeogenic enzymes. “GID” is a collection of E3 ligase complexes; a core scaffold, RING-type catalytic core and supramolecular module along with interchangeable substrate receptors select targets for ubiquitylation. However, knowledge of additional cellular factors directly regulating GID-type E3s remains rudimentary. Here, we structurally and biochemically characterize Gid12 as a modulator of the GID E3 ligase complex targeting Fbp1. Our collection of cryo-EM reconstructions shows that Gid12 forms an extensive interface sealing the substrate receptor Gid4 onto the scaffold, and remodeling the degron binding site. Gid12 also sterically blocks a recruited Fbp1 from the ubiquitylation active sites. Our analysis of the role of Gid12 establishes principles that may more generally underlie E3 ligase regulation.

INSTRUMENT(S): timsTOF Pro, Orbitrap Exploris 480

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Barbara Steigenberger

LAB HEAD: Brenda A. Schulman

PROVIDER: PXD028579 | Pride | 2022-05-29

REPOSITORIES: Pride

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			Action	DRS
	20200923_TTOF01_LC12_17_SA_Orbi3661_Sample01_C1_1_2465.d.7z	Other
	20200923_TTOF01_LC12_17_SA_Orbi3661_Sample02_C2_1_2466.d.7z	Other
	20200923_TTOF01_LC12_17_SA_Orbi3661_Sample03_C3_1_2467.d.7z	Other
	20200923_TTOF01_LC12_17_SA_Orbi3661_Sample04_C4_1_2469.d.7z	Other
	20200923_TTOF01_LC12_17_SA_Orbi3661_Sample05_C5_1_2470.d.7z	Other

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