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.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 185331 | pGEX-6P-1 AUP1 292-410 | Bacterial expression of AUP1 with GST fusion protein for binding assays |
| 185332 | pGEX-6P-1 AUP1 292-410 5E | Bacterial expression of AUP1 with GST fusion protein for binding assays |
| 185333 | pGEX-6P-1 AUP1 292-410 A397D | Bacterial expression of AUP1 with GST fusion protein for binding assays |
| 185334 | pGEX-6P-1 AUP1 61-410 | Bacterial expression of AUP1 with GST fusion protein for binding assays |
| 185335 | pETDUET-GST-AUP1 379-410:Ube2G2 | Used for co-expression of the AUP1 G2BR with UBE2G2 for crystallography |
| 185336 | pCDNA3-AUP1-FLAG | Mammalian expression of AUP1 |
| 185337 | pCDNA3-AUP1-FLAG DTM | Assessing domain requirements of AUP1 |
| 185339 | pCDNA3-AUP1-FLAG-DCUE | Assessing domain requirements of AUP1 |
| 185340 | pCDNA3-AUP1-FLAG-DATDCUE | Assessing domain requirement sof AUP1 |
| 185341 | pCDNA3-AUP1-FLAG 5E | Assessing G2BR function in AUP1 |
| 185342 | pCDNA3-AUP1 A397D | Assessing G2BR function in AUP1 |
| 185343 | pEGFP-C1-G2BRAUP1 | Expression of soluble G2BR, amino acids 378-404 of human AUP1 |
| 185344 | pEGFP-C1-G2BRgp78 | Expression of soluble G2BR, amino acids 575-600 of human gp78 |
| 185345 | pEGFP-C1-G2BRSCR | Expression of soluble G2BR with scrambled sequence |
| 185346 | pcDNA3.1(+)-Cue1pTM-MYC-UBE2G2 | Membrane anchoring of UBE2G2. Encodes transmembrane domain of S. Cerevisiae Cue1p fused to MYC-UBE2G2 |
| 185347 | pcDNA3.1(+)-Cue1pTM-MYC-UBE2G2-FLAG | Membrane anchoring of UBE2G2 and co-IP. Encodes transmembrane domain of S. Cerevisiae Cue1p fused to MYC-UBE2G2 with C-terminal FLAG tag |
| 185348 | pET3a-UBE2J1 1-283 | In vitro transcription and translation in bacterial system |
| 185349 | pGEX-6P-1-HRD1 aa 236-444 | RING domain-containing GST fusion protein for ubiquitination assays. |
| 185350 | pcDNA4-NHK-MYC-His6 | Assessing of ubiquitination in cells |
| 185351 | pcDNA4-RI1-332-MYC-His6 | Mammalian expression of Ribophorin I |
| 185352 | pGEX-4T-3-TRC8 513-664 | RING domain-containing GST fusion protein for ubiquitination assays. |
| 185353 | pGEX-6P-1-gp78 RING | RING domain-containing GST fusion protein for ubiquitination assays. |
| 185354 | pGEX-6P-1-gp78 429-611 | GST fusion protein for quantification of gp78 levels |
| 185355 | pCINeo-gp78 Mutant G2BR | Assessing cellular requirement for G2BRgp78 |
| 185356 | pGEX-6P-1-Cys-HA-Ubiquitin | In vitro ubiquitination assays |