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A structurally conserved site in AUP1 binds the E2 enzyme UBE2G2 and is essential for ER-associated degradation.
Smith CE, Tsai YC, Liang YH, Khago D, Mariano J, Li J, Tarasov SG, Gergel E, Tsai B, Villaneuva M, Clapp ME, Magidson V, Chari R, Byrd RA, Ji X, Weissman AM
PLoS Biol. 2021 Dec 8;19(12):e3001474. doi: 10.1371/journal.pbio.3001474. eCollection 2021 Dec.
PubMed Article

Plasmids from Article

ID Plasmid Purpose
185331pGEX-6P-1 AUP1 292-410Bacterial expression of AUP1 with GST fusion protein for binding assays
185332pGEX-6P-1 AUP1 292-410 5EBacterial expression of AUP1 with GST fusion protein for binding assays
185333pGEX-6P-1 AUP1 292-410 A397DBacterial expression of AUP1 with GST fusion protein for binding assays
185334pGEX-6P-1 AUP1 61-410Bacterial expression of AUP1 with GST fusion protein for binding assays
185335pETDUET-GST-AUP1 379-410:Ube2G2Used for co-expression of the AUP1 G2BR with UBE2G2 for crystallography
185336pCDNA3-AUP1-FLAGMammalian expression of AUP1
185337pCDNA3-AUP1-FLAG DTMAssessing domain requirements of AUP1
185339pCDNA3-AUP1-FLAG-DCUEAssessing domain requirements of AUP1
185341pCDNA3-AUP1-FLAG 5EAssessing G2BR function in AUP1
185342pCDNA3-AUP1 A397DAssessing G2BR function in AUP1
185343pEGFP-C1-G2BRAUP1Expression of soluble G2BR, amino acids 378-404 of human AUP1
185344pEGFP-C1-G2BRgp78Expression of soluble G2BR, amino acids 575-600 of human gp78
185345pEGFP-C1-G2BRSCRExpression of soluble G2BR with scrambled sequence
185346pcDNA3.1(+)-Cue1pTM-MYC-UBE2G2Membrane anchoring of UBE2G2. Encodes transmembrane domain of S. Cerevisiae Cue1p fused to MYC-UBE2G2
185347pcDNA3.1(+)-Cue1pTM-MYC-UBE2G2-FLAGMembrane anchoring of UBE2G2 and co-IP. Encodes transmembrane domain of S. Cerevisiae Cue1p fused to MYC-UBE2G2 with C-terminal FLAG tag
185348pET3a-UBE2J1 1-283In vitro transcription and translation in bacterial system
185349pGEX-6P-1-HRD1 aa 236-444RING domain-containing GST fusion protein for ubiquitination assays.
185350pcDNA4-NHK-MYC-His6Assessing of ubiquitination in cells
185351pcDNA4-RI1-332-MYC-His6Mammalian expression of Ribophorin I
185352pGEX-4T-3-TRC8 513-664RING domain-containing GST fusion protein for ubiquitination assays.
185353pGEX-6P-1-gp78 RINGRING domain-containing GST fusion protein for ubiquitination assays.
185354pGEX-6P-1-gp78 429-611GST fusion protein for quantification of gp78 levels
185355pCINeo-gp78 Mutant G2BRAssessing cellular requirement for G2BRgp78
185356pGEX-6P-1-Cys-HA-UbiquitinIn vitro ubiquitination assays

Antibodies from Article