Table I.

A yeast two-hybrid assay

ROKβ regions boundROKα regions bound
Gem full-length (2–296)a, c, e
Gem Δc (2–262)a, c, e
Gem core (71–262)a, c, e
Gem(S89N)a, c, e
Rad (41–308)c
Rad Δc (41–274)a, c, e
Rem1 full-length (2–297)
Rem2 full-length (2–272)
Rem1 Δc (2–254)
Rho Q63La, ca, c
  • Y190 yeast were transformed with combinations of plasmids expressing Gal4 DNA binding domain–GTP binding protein and Gal4 activation domain–ROKα or ROKβ fragments. The letters refer to the diagrammatic representation of the various ROK fragments shown in Fig. 1. 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.