Table 4.
Pex13-SH3 mutants have lost two-hybrid interaction with Pex14p but not with Pex5p
| GAL4 AD fusion | GAL4 DB fusion | β-Galactosidase activity (RLU/mg protein ± SD) | Growth on his− |
|---|---|---|---|
| — | — | 1.0 ± 0.1 | - |
| Pex5p | — | 0.5 ± 0.1 | - |
| Pex5p | Pex13-SH3 | 3,000 ± 900 | ++ |
| Pex5p | Pex13-SH3(E320K) | 3,500 ± 400 | ++ |
| Pex5p | Pex13-SH3(W349A) | 1,600 ± 800 | ND |
| — | Pex13-SH3 | 3.8 ± 1.0 | - |
| — | Pex13-SH3(E320K) | 3.1 ± 0.2 | - |
| — | Pex13-SH3(W349A) | 1.3 ± 0.1 | - |
| Pex14p | — | 0.8 ± 0.1 | - |
| Pex14p | Pex13-SH3 | 39 ± 12 | ++ |
| Pex14p | Pex13-SH3(E320K) | 2.7 ± 0.2 | - |
| Pex14p | Pex13-SH3(W349A) | 2.0 ± 0.5 | - |
Plasmids encoding GAL4 DB and GAL4 AD fusions as indicated were transformed to two-hybrid yeast strains PCY2 and HF7c. Two-hybrid interaction was quantitated in PCY2 by measuring the β-galactosidase activity as described in MATERIALS AND METHODS. Indicated numbers are the mean of two independent measurements in triplicate cultures ± SD. Two-hybrid interaction was measured in HF7c by growth in the absence of histidine (++, growth; -, no growth; ND, not determined). RLU, relative light units.