TABLE 1.
Two-hybrid interaction of Rho3 with Exo70 and Myo2a
| Construct | β-Galactosidase activity (Miller units; mean ± SD)
|
||
|---|---|---|---|
| Exo70 | RBP4 | RBP26 | |
| Rho3E129 | 3.28 ± 0.10 | 48.09 ± 12.7 | 3.48 ± 0.83 |
| Rho3E129/A48 | 0.20 ± 0.02 | 0.17 ± 0.04 | 0.11 ± 0.02 |
| Rho3E129/S47 | 0.26 ± 0.02 | 6.79 ± 1.13 | 0.35 ± 0.18 |
| Rho3E129/N30 | 0.20 ± 0.03 | 0.23 ± 0.01 | 0.19 ± 0.03 |
a
The liquid β-galactosidase assay was carried out as described by Poullet and Tamanoi (44). RHO3 was fused to the DNA binding domain vector pGBT9. EXO70, RBP4, and RBP26 were fused to the activation domain vector pGAD-GH or pGAD424. The interaction between pGBT9 and pGAD-GH (0.19 ± 0.04 U) was used as a negative control, and the interaction between pGBT9-NF1 and pGAD-HrasV12 (activated Hras) (4.35 ± 2.11) was used as a positive control as previously described (44). These experiments were repeated three times with similar results.