Interconversion between Anticipatory and Active GID E3 Ubiquitin Ligase Conformations via Metabolically Driven Substrate Receptor Assembly

• Published in: Molecular cell Vol. 77; no. 1; pp. 150 - 163.e9
• Main Authors: Qiao, Shuai, Langlois, Christine R., Chrustowicz, Jakub, Sherpa, Dawafuti, Karayel, Ozge, Hansen, Fynn M., Beier, Viola, von Gronau, Susanne, Bollschweiler, Daniel, Schäfer, Tillman, Alpi, Arno F., Mann, Matthias, Prabu, J. Rajan, Schulman, Brenda A.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 02.01.2020
• Subjects: active sites; Animals; biochemical pathways; carbon; Catalytic Domain - physiology; Cell Line; cryo-electron microscopy; cryo-EM; Cryoelectron Microscopy - methods; CTLH; E3 ligase; genetics; GID; gluconeogenesis; Gluconeogenesis - physiology; glucose; Glucose - metabolism; homeostasis; Humans; Lysine - metabolism; metabolism; N-degron; N-end rule; receptors; Saccharomyces cerevisiae; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - metabolism; stress response; ubiquitin; ubiquitin-protein ligase; Ubiquitin-Protein Ligases - metabolism; Ubiquitination - physiology; CTLH; N-degron; E3 ligase; stress response; GID; N-end rule; metabolism; cryo-EM; gluconeogenesis; ubiquitin
• ISSN: 1097-2765; 1097-4164; 1097-4164
• DOI: 10.1016/j.molcel.2019.10.009

Cells respond to environmental changes by toggling metabolic pathways, preparing for homeostasis, and anticipating future stresses. For example, in Saccharomyces cerevisiae, carbon stress-induced gluconeogenesis is terminated upon glucose availability, a process that involves the multiprotein E3 ligase GIDSR4 recruiting N termini and catalyzing ubiquitylation of gluconeogenic enzymes. Here, genetics, biochemistry, and cryoelectron microscopy define molecular underpinnings of glucose-induced degradation. Unexpectedly, carbon stress induces an inactive anticipatory complex (GIDAnt), which awaits a glucose-induced substrate receptor to form the active GIDSR4. Meanwhile, other environmental perturbations elicit production of an alternative substrate receptor assembling into a related E3 ligase complex. The intricate structure of GIDAnt enables anticipating and ultimately binding various N-degron-targeting (i.e., “N-end rule”) substrate receptors, while the GIDSR4 E3 forms a clamp-like structure juxtaposing substrate lysines with the ubiquitylation active site. The data reveal evolutionarily conserved GID complexes as a family of multisubunit E3 ubiquitin ligases responsive to extracellular stimuli. [Display omitted] •Genetics, biochemistry, proteomics, and cryo-EM define GID E3 ligase regulation•Carbon stress induces assembly of an inactive anticipatory GIDAnt complex•Environmental perturbations trigger substrate receptor assembly into active GID E3s•Structural model of N-degron substrate ubiquitylation by multisubunit RING-RING E3 Stunning structural intricacies of the yeast GID system, now revealed more than 25 years since the discovery of glucose-induced degradation and of a protein’s N terminus serving as a degron, provide a blueprint for a family of multisubunit E3s regulated through assembly with interchangeable substrate receptors induced by distinct environmental perturbations.