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. 2009 Dec 2;106(50):21419–21424. doi: 10.1073/pnas.0910601106

Fig. 4.

Fig. 4.

OST1 phosphorylates SLAC1 N-terminal and C-terminal domains and is inactivated by PP2CA. (A) OST1 interacted with PP2CA and several other PP2C members. OST1 was cloned into the pGBTKT7 and the PP2Cs were in pGAD.GH. Yeast cells were as described in Materials and Methods and in Fig. 1. AIPH1: AIP1 homologue 1 (At2g29380); AIPH2: AIP1 homologue 2 (At5g59220). (B) Phosphorylation of SLAC1 by OST1. The contents of the kinase assays were shown at the top of the autoradiography picture. The 32P-labeled protein bands are indicated by arrows and names of the proteins at the left side. (C) PP2CA and PP2CA null mutant inhibit OST1 autokinase activity and activity against SLAC1. The assays are presented in the same manner as described in (B). (D) OST1 interacted with PP2CA and PP2CA null mutant in a yeast two hybrid assay described in (A). The PP2CA null mutant, as well as PP2CA, was in pGAD.GH. (E) In vitro dephosphorylation assay of PP2CA and PP2CA null mutant (see Materials and Methods)