Table 3.
Smy1p interacts with Myo2p in two different two-hybrid systems, independent of vectors used
| 2-Hybrid interaction | |
|---|---|
| LexA System constructsa | |
| pBTM-SMY1b + pACT-MYO2d | ++d |
| pBTM-SMY1 + Myo2R (pGAD)e | ++ |
| pBTM-MYO2 + pACT-SMY1 | ++ |
| pBTM-MYO2 + pGAD-SMY1e | ++ |
| Gal4 system constructsa | |
| pAS1-SMY1 + pACT-MYO2 | ++ |
| pAS1-SMY1 + pGAD-MYO2 | − |
All fusion proteins were detected by Western analysis.
The N terminus of all Smy1p fusions is truncated by 18 amino acids, except for pBTM-SMY1, which is truncated by 43 amino acids.
The Myo2p fusions begin with the coiled-coil domain (amino acid 927) and continue to the C terminus of the tail.
++, robust two-hybrid interaction which corresponds to 15-fold greater activity than the background activity determined with the individual plasmids; −, no two-hybrid interaction detected.
These pairs were tested in a liquid beta-galactosidase assay (Kaiser et al., 1994) and found to have 15-fold greater activity than the background activity determined with the individual plasmids.