Figure 6.
SCaBP8 Phosphorylation Affects Its Interaction with SOS2.
(A) The SCaBP8S237A mutation reduces SCaBP8 interaction with SOS2 in yeast. Yeast strain AH109 harboring SOS2 in the pGADT7 vector was transformed with SCaBP8, SCaBP8S237D, or SCaBP8S237A in the pGBKT7 vector. Growth of the transformed yeast was assayed on media minus Trp and Leu (top panel), minus Trp, Leu, and His (middle panel), or minus Trp, Leu, and His with 2.5 mM 3-amino-1,2,4-triazole (bottom panel).
(B) The SCaBP8S237A mutation reduces SCaBP8 interaction with SOS2 in Arabidopsis. Flag-SCaBP8, flag-SCaBP8S237D, and flag-SCaBP8S237A were expressed in protoplasts from myc-SOS2 transgenic plants. Total protein was analyzed using anti-myc and anti-flag antibodies (Input). After immunoprecipitation with anti-myc conjugated agarose, the components in the pellet were detected via immunoblot analysis (Output). SC8, SCaBP8; SD, SCaBP8S237D; SA, SCaBP8S237A.