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A structurally conserved site in AUP1 binds the E2 enzyme UBE2G2 and is essential for ER-associated degradation
- Source :
- PLoS Biology, Vol 19, Iss 12, p e3001474 (2021), PLoS Biology
- Publication Year :
- 2021
- Publisher :
- Public Library of Science (PLoS), 2021.
-
Abstract
- Endoplasmic reticulum–associated degradation (ERAD) is a protein quality control pathway of fundamental importance to cellular homeostasis. Although multiple ERAD pathways exist for targeting topologically distinct substrates, all pathways require substrate ubiquitination. Here, we characterize a key role for the UBE2G2 Binding Region (G2BR) of the ERAD accessory protein ancient ubiquitous protein 1 (AUP1) in ERAD pathways. This 27-amino acid (aa) region of AUP1 binds with high specificity and low nanomolar affinity to the backside of the ERAD ubiquitin-conjugating enzyme (E2) UBE2G2. The structure of the AUP1 G2BR (G2BRAUP1) in complex with UBE2G2 reveals an interface that includes a network of salt bridges, hydrogen bonds, and hydrophobic interactions essential for AUP1 function in cells. The G2BRAUP1 shares significant structural conservation with the G2BR found in the E3 ubiquitin ligase gp78 and in vitro can similarly allosterically activate ubiquitination in conjunction with ERAD E3s. In cells, AUP1 is uniquely required to maintain normal levels of UBE2G2; this is due to G2BRAUP1 binding to the E2 and preventing its rapid degradation. In addition, the G2BRAUP1 is required for both ER membrane recruitment of UBE2G2 and for its activation at the ER membrane. Thus, by binding to the backside of a critical ERAD E2, G2BRAUP1 plays multiple critical roles in ERAD.<br />This study shows that a 27-amino acid domain that binds the ubiquitin conjugating enzyme UBE2G2, and which is found in two different proteins (AUP1 and gp78), is critical for endoplasmic reticulum-associated degradation (ERAD).
- Subjects :
- QH301-705.5
Artificial Gene Amplification and Extension
macromolecular substances
Transfection
Research and Analysis Methods
Endoplasmic Reticulum
Biochemistry
Polymerase Chain Reaction
General Biochemistry, Genetics and Molecular Biology
Protein Domains
Cell Line, Tumor
Solid State Physics
Humans
Amino Acid Sequence
Post-Translational Modification
Biology (General)
Molecular Biology Techniques
Molecular Biology
Secretory Pathway
Crystallography
General Immunology and Microbiology
Physics
General Neuroscience
Ubiquitination
Organisms
Fungi
Biology and Life Sciences
Proteins
Eukaryota
Membrane Proteins
Cell Biology
Endoplasmic Reticulum-Associated Degradation
Condensed Matter Physics
Lipids
Yeast
Cell Processes
Physical Sciences
Ubiquitin-Conjugating Enzymes
Crystal Structure
Cell lines
Cellular Structures and Organelles
Biological cultures
HT1080 cells
General Agricultural and Biological Sciences
Research Article
Protein Binding
Subjects
Details
- ISSN :
- 15457885
- Volume :
- 19
- Database :
- OpenAIRE
- Journal :
- PLOS Biology
- Accession number :
- edsair.doi.dedup.....0901fbce041c16d2a2ac55caf44214d1