Skip to main content
. 2016 May 27;473(11):1579–1591. doi: 10.1042/BCJ20160085

Figure 2. Investigation of the nature of the interaction mediated by E596 in V617F activation.

Figure 2

(A) STAT5 transcriptional activity was measured in γ2A cells by luciferase assay. Mutation of E596 to either arginine or lysine induces a decrease in constitutive activity of JAK2 V617F up to 71% and 60% respectively whereas substitution to aspartate increases the activity of the mutant protein up to 86%. The activity of the wild-type JAK2 amounted to 25% of the activity of JAK2 V617F. Shown are means for three independent experiments each performed in triplicate and data were normalized by taking 100% as the basal activity of JAK2 V617F. (B) Substitution of E596 by protonated residues in JAK2 V617F displays an Epo-induced response similar to JAK2 WT. These two mutants behave like the wild-type JAK2. Shown are means±S.E.M. for three independent experiments done in triplicate. n.s, not significant. (C) BaF3-EpoR stably expressing JAK2 double mutants were starved for 4 h and allow to grow in a cytokine-free media. The number of living cells was evaluated by ATP measurement during 6 days. Only JAK2 V617F and JAK2 V617F/E596D transform the BaF3 into factor-independent cells. Although E596/K and A mutations prevent autonomous grow of JAK2 V617F.