Table 3.
Two-hybrid analysis
| STE4 | STE18 | STE50 | STE20 | STE5 | STE11 | STE7 | FUS3 | KSS1 | STE12 | |
|---|---|---|---|---|---|---|---|---|---|---|
| pep1(I) | − | − | − | − | − | − | − | − | − | − |
| pep2(I) | − | − | − | − | 9 | − | − | − | − | − |
| pep3(C) | − | − | − | − | − | − | − | − | − | − |
| pep4(C) | − | − | − | − | − | − | − | − | − | − |
| pep5(C) | − | − | − | − | − | − | − | − | − | − |
| orf1 | − | − | − | − | + | + | − | − | − | − |
| orf2 | − | − | - | − | 134 | 17 | − | − | − | − |
| orf3 | − | − | − | − | 13 | − | − | − | − | − |
| orf4 | − | − | − | − | 19 | − | − | − | − | − |
| orf5 | − | − | − | − | − | 9 | − | − | − | − |
| orf6 | − | − | − | − | − | − | − | − | − | − |
| orf7 | − | − | − | − | − | − | − | − | − | − |
| orf8 | − | − | − | − | − | − | − | − | − | − |
| orf9 | − | − | − | − | 14 | − | − | − | − | − |
| orf10 | − | 18 | − | − | − | − | − | − | − | − |
| orf11 | − | − | − | − | − | − | − | − | − | − |
Shown are the results of pairwise examinations of each perturbagen with various known members of the pheromone response pathway. Numbers given for each positive interaction represent units of β-galactosidase activity (micromoles of o-nitrophenyl-β-d-galactopyranoside hydrolysed min−1 cell−1) determined in quantitative liquid assays according to the methods described (ref. 22 and Materials and Methods). Units of activity were standardized to the level of activity measured for each binding domain partner combined with an activation domain SV40-large-T antigen fusion (standardized to 1 unit). In all cases, units of activity were determined from two independent pairings, each measured in triplicate. The average SD was <20% with a maximum of 41% (the orf9/STE5 pairing). In all cases except orf10, the numbers shown were determined from the orf-AD fusion/STE-BD fusion pairing. The positive interactions marked with a + for orf1 were not observed in this study and are extrapolated from ref. 20.