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p54/46 ATF-2 kinases were bound to the GST-ATF-2 column and after elution of the kinases from the column with 2 M urea, fractions were tested for kinase activity (Figure 7A)....we used a mutant of ATF-2 containing point mutations at positions Thr69, Thr71 and Ser9O. As shown in Figure 7B (right panel), both the kinases purified with GST-ATF-2 and with GST-Jun (JNK) hardly phosphorylated GST-ATF-2 protein lacking the putative phosphoacceptor sites (M).....The GST-c-Jun protein containing point mutations at positions Ser63 and Ser73 (M) was phosphorylated less efficiently than wild-type c-Jun (W) by ATF-2 kinases (Figure 7B, left panel), similar to that found for p54/46 JNK (Hibi et al., 1993).
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