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. 2018 Mar 23;15(6):739–755. doi: 10.1080/15476286.2018.1445958

Figure 2.

Figure 2.

Identification of the PABP1 phosphorylation motifs. (A) Structural domain organization of the L. infantum PABP1 homologue. The protein's RRM region consisting of the first two thirds of PABP1 (encompassing all four RRMs) plus the “Linker” segment and the “MLLE” domain are shown. Seven putative phosphorylation sites are indicated, six of which were targeted by site-directed mutagenesis (circled). Also highlighted are the three sets of amino acid triplets chosen for mutagenesis and localized between RRMs 1 and 2 (LMW), within RRM2 (YGF) or within the MLLE domain (TGM) The N-terminus and C-terminus fragments used for the pull-down assays are also indicated. (B) Expression of HA-tagged, episomally encoded, PABP1 evaluated with a monoclonal commercial anti-HA antibody. The left panel compared the expression of wild-type PABP1 (HA-PABP1) during three representative stages of the parasite growth curve. The right panel evaluates the expression, under the same conditions of the PABP1 mutant (TP-SP Mut) where all six putative phosphorylation sites were targeted by site directed mutagenesis.