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. 2022 Sep 20;298(11):102513. doi: 10.1016/j.jbc.2022.102513

Figure 2.

Figure 2

Isoform-specific residues of the 14-3-3 proteins promote interaction with UL97.A, schematic diagram of the sixth, seventh, and eighth α helices from seven 14-3-3 isoforms. The asterisks (∗) indicate the amino acid residues tested for binding specificity with UL97. B, U-2 OS cells were transfected with an expression vector for HA-tagged UL97 together with expression vectors for either WT 14-3-3σ or a mutant (M) with three amino acid changes (A147S/H180Y/I191C). Lysates (Input) were subjected to immunoprecipitation (IP) with anti-HA antibody (IP: anti-HA) and analyzed by Western blot with the indicated antibodies. C, IPs as in panel (B) with triple amino acid mutants (M) of 14-3-3β (S147A/Y180H/C191I), 14-3-3ε (S148A/Y181H/C192I), 14-3-3η (S150A/Y183H/C194I), 14-3-3γ (S150A/Y183H/C194I), or 14-3-3θ (S145A/Y178H/C189I). More expression vector for mutated 14-3-3β (1 μg) than WT 14-3-3β (0.4 μg) was used to obtain similar expression levels. D, IPs as in panel (B) with a 14-3-3ζ (A134C) mutant (Mut C). E, IPs as in panel (B) with a 14-3-3θ (C134A) mutant (Mut A). For all panels, representative images from one of two independent biological replicates are shown.