Figure 1
L42R redistributes PTENA4 and ePTEN to the nucleus from the plasma membrane.
(A) Associations between the core region (e.g. the phosphatase and C2 domains) and the tail of the same PTEN molecule (closed conformation) are maintained through the phosphorylation of the tail (indicated by “P”). In contrast, the core region of PTENA4 dissociates from the tail (open conformation) and binds to an exogenously added tail domain22. ePTEN exists in an open conformation due to mutations in the core region (asterisks). Therefore, ePTEN cannot bind to the exogenously added tail22. (B) Dictyostelium cells expressing the indicated forms of PTEN-GFP were observed by fluorescence microscopy. Bar, 10 μm. (C) Intensity of GFP at the plasma membrane and in the nucleus was quantified relative to that in the cytosol. Values represent the mean ± SD (n ≥ 8). (D) HEK293 cells expressing the indicated forms of PTEN-GFP were observed by fluorescence microscopy. Bar, 10 μm. (E) Whole-cell lysates from Dictyostelium expressing the indicated PTEN-GFP proteins were incubated with PTEN352–403-YFP-FLAG expressed in HEK293 cells. PTEN352–403-YFP-FLAG was immunoprecipitated with beads coupled to anti-FLAG antibodies. (F) The indicated PTEN-GFP proteins were immunopurified from Dictyostelium cells (Fig. S2) and phosphatase activities were measured. Values represent the mean ± SD (n = 4).
