Structural insights into E1 recognition and the ubiquitin-conjugating activity of the E2 enzyme Cdc34
Year: 2019
Type: article
Abstract: Ubiquitin (Ub) signaling requires the sequential interactions and activities of three enzymes, E1, E2, and E3. Cdc34 is an E2 that plays a key role in regulating cell cycle progression and requires unique structural elements to function. The molecular basis by which Cdc34 engages its E1 and the structural mechanisms by which its unique C-terminal extension functions in Cdc34 activity are unknown. Here, we present crystal structures of Cdc34 alone and in complex with E1, and a Cdc34~Ub thioester mimetic that represents the product of Uba1-Cdc34 Ub transthiolation. These structures reveal conformational changes in Uba1 and Cdc34 and a unique binding mode that are required for transthiolation. The Cdc34~Ub structure reveals contacts between the Cdc34 C-terminal extension and Ub that stabilize Cdc34~Ub in a closed conformation and are critical for Ub discharge. Altogether, our structural, biochemical, and cell-based studies provide insights into the molecular mechanisms by which Cdc34 function in cells.
Source: Nature Communications
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $5,200
Funders: U.S. Department of Energy, U.S. Department of Health and Human Services, Abney Foundation, Medical University of South Carolina, University of South Carolina, National Institutes of Health, Office of Science, National Heart, Lung, and Blood Institute, National Cancer Institute, National Institute of General Medical Sciences, National Institute of Dental and Craniofacial Research, National Center for Research Resources, Office of Research Infrastructure Programs, National Institutes of Health, Argonne National Laboratory
Awards: G102163711, G107546177, G1183574029, G1188805541, G1313983767, G1505794299, Mechanisms of Esophageal Carcinogenesis, G1664757510, G1785301001, G204618756, G2101401529, G2164036156, G2287611682, G254657075, G262962599, G2887060908, G2894532024, G3067649710, G3075337988, Pixel Array Detector for X-ray Crystallography, G3337633282, G340565428, G3407338877, G3549828030, Crystallization robotics to support X-ray crystallography at MUSC, G3754226247, G3869404228, G409777406, Nuclear Accumulation of D-type Cyclins and Oncogenesis, G4557230613, G4656156008, G498139845, G5106849484, Structural Biology of the Ubiquitin Conjugation System, G5201631574, G5384392517, G5520316217, G5607048534, G6106915579, G620160007, G666778509, G6848031779, G6918803902, G7257490191, Investigation of Cdc34 Molecular Recognition by its Enzyme Partners, Uba1 and SCF Ligases., G7368700025, NE-CAT Center for Advanced Macromolecular Crystallography, G7756544684, G8158403594, Integrative Training in Oncogenic Signaling, G858945595, G9000038451, G9000072012, G9000119994, G9000195902, G9000263424, T-COHR: Training in Craniofacial and Oral Health Research, Structure and Function of the Essential Cell Cycle Regulator Cdc34, G969889393, G993149817
OpenAlex ID: W2962948444