Table 1. Various functional roles of CBP-interacting sites I and II of GRIP1 with GR, PR and TR in HeLa, HEK-293 and CV-1 cell lines.
HeLa, HEK-293 and CV-1 cells were transfected with plasmids encoding NR (0.1 μg, except for 0.04 μg of TR in HeLa cells) and the GRIP1 fragment indicated (0.35 μg), which also contained the LUC reporter gene (0.25 μg), as shown. Transfected cultures were grown with the appropriate hormone (100 nM dexamethasone for GR; 10 nM progesterone for PR; 100 nM 3,3′,5-tri-iodothyronine for TR). LUC fold activation is expressed relative to that of samples containing NR, but no GRIP1. Similar activity profiles were observed in three independent experiments.
| Fold activation | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| CBP-interacting site status | GR+MMTV-LUC | PR+MMTV-LUC | TR+MMTV(TRE)-LUC | ||||||||
| GRIP1 | I | II | HeLa | HEK-293 | CV-1 | HeLa | HEK-293 | CV-1 | HeLa* | HEK-293 | CV-1 |
| Vector | − | − | 1±0.2 | 1±0.1 | 1±0.2 | 1±0.2 | 1±0.1 | 1±0.1 | 1±0.1 | 1±0.2 | 1±0.01 |
| 5–1462 | + | + | 10±1 | 1±0.1 | 32±1 | 5±0.6 | 19±2 | 12±0.2 | 17±2 | 5±0.3 | 5±0.7 |
| 5–1462 | − | + | 6±0.7 | 1±0.1 | 18±1 | 2±0.5 | 5±0.4 | 3±0.3 | 2±0.3 | 0.5±0.05 | 0.5±0.02 |
| 5–1462 | + | − | 13±1 | 1±0.1 | 24±0.2 | 5±0.5 | 29±0.2 | 13±1 | 44±8 | 6±0.6 | 3±0.7 |
| 563–1121 | + | + | 7±1 | 0.6±0.01 | 48±5 | 2±0.1 | 6±0.1 | 7±0.3 | 12±1 | 7±0.6 | 13±0.5 |
| 563–1121 | − | + | 6±0.1 | 1±0.01 | 34±1 | 1±0.5 | 6±0.4 | 5±0.2 | 1±0.1 | 0.3±0.01 | 0.7±0.09 |
| 563–1121 | + | − | 7±0.7 | 0.8±0.01 | 31±0.6 | 2±0.7 | 5±0.3 | 6±0.9 | 6±0.1 | 6±0.3 | 4±0.2 |
| 563–1094 | + | − | 9±0.4 | 1±0.05 | 36±1 | 2±0.1 | 7±0.4 | 8±1 | 6±0.2 | 6±0.2 | 4±0.4 |
| 563–1012 | − | − | 7±0.2 | 1±0.02 | 31±6 | 2±0.1 | 1±0.1 | 8±1 | 2±0.3 | 0.6±0.05 | 1±0.2 |
* Fold values are calculated based on the relative data shown in Figure 5(a).