Table 1.
5LO interacts specifically with CLP, partial TRAP-1 (ΔTRAP-1), and the partial K12H4.8 (ΔK12H4.8) homologue in the yeast two-hybrid system
| Gal4 DNA-BD constructs | Gal4 DNA-AD constructs | Reporter genes
|
||
|---|---|---|---|---|
| Adenine | Histidine | lacZ | ||
| Empty | Empty | − | − | White |
| SNF1 | Empty | − | − | White |
| Empty | SNF4 | − | − | White |
| SNF1 | SNF4 | + | + | Blue |
| 5LO | Empty | − | − | White |
| 5LO | SNF4 | − | − | White |
| 5LO | SV40 large T-antigen | − | − | White |
| Empty | CLP | − | − | White |
| SNF1 | CLP | − | − | White |
| MGST-I | CLP | − | − | White |
| 5LO | CLP | + | + | Blue |
| Empty | ΔTRAP-1 | − | − | White |
| SNF1 | ΔTRAP-1 | − | − | White |
| MGST-I | ΔTRAP-1 | − | − | White |
| 5LO | ΔTRAP-1 | + | + | Blue |
| Empty | ΔK12H4.8 homologue | − | − | White |
| SNF1 | ΔK12H4.8 homologue | − | − | White |
| MGST-I | ΔK12H4.8 homologue | − | − | White |
| 5LO | ΔK12H4.8 homologue | + | + | Blue |
+, growth; −, no growth. The plasmids expressing the Gal4 fusion proteins were cotransformed into the yeast strain PJ69-4A, and transformants were selected on SD/−Leu/−Trp. Three or four independent colonies were streaked on SD/−Leu/−Trp/−Ade, on SD/−Leu/−Trp/−His + 4 mM 3-amino-1,2,4-triazole, and on SD/−Leu/−Trp + 5-bromo-4-chloro-3-indolyl β-d-galactoside plates to test for the adenine, histidine, and lacZ reporter genes respectively, and incubated at 30°C for 3–5 days. Growth and color of the colonies then were determined.