Table II.

A yeast two-hybrid assay

Gem coreRad ΔcRho Q63L
ROKβ (787–1027)+++
ROKβ (787–1027) [N1004T, K1005T]++
ROKβ (787–976)++
ROKβ (907–1027)+
ROKβ (726–1027, Δ787–906)+
  • Y190 yeast were transformed with combinations of plasmids expressing Gal4 DNA binding domain–GTP binding protein and Gal4 activation domain–ROKβ fragments. ROKβ (787–976) is adjacent to but does not include the p21 binding domain. ROKβ (907–1027) contains the previously defined p21 binding domain. At least three independent transformants were assayed in each case. Interactions were considered positive if growth occurred after 3 d at 30°C on selective medium lacking histidine and containing at least 2 mM aminotriazole and if a blue color was observed within 5 min of initiating an in situ β-galactosidase assay.