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. 2019 Aug 5;15(8):e1007962. doi: 10.1371/journal.ppat.1007962

Fig 6. Analysis of the membrane binding site of ACBD3 in complex with the enterovirus 3A protein.

Fig 6

a, Membrane binding model of the GOLD: poliovirus 3A complex. The ACBD3 GOLD domain is shown in cartoon representation with a semi-transparent surface and colored in gold except for the membrane binding site (MBS) composed of R399, L514, W515, and R516, which is colored in green. The poliovirus 3A protein is depicted in blue. b, Localization of the ACBD3 mutants. EGFP-fused wild-type ACBD3 or its mutants were overexpressed in HeLa ACBD3 KO cells. Cells were fixed and immunostained with the anti-giantin antibody (marker of Golgi). Scale bars represent 10 μm. c, Localization of the ACBD3 mutants in 3A-expressing cells. EGFP-fused ACBD3 mutants were co-expressed with myc-tagged CVB3 3A in HeLa ACBD3 KO cells. Cells were fixed and immunostained with the anti-myc and anti-giantin (marker of Golgi) antibodies. Scale bars represent 10 μm. d, Statistical analysis of the ACBD3-giantin co-localization from (b) and (c) is presented as Mander's correlation coefficients ± SDs from at least 12 cells from 2 independent experiments. e, Rescue of enterovirus replication by the ACBD3 mutants. HeLa ACBD3 KO cells were transfected with wild-type ACBD3 or its mutants, and enterovirus replication was determined using the Renilla luciferase-expressing CVB3 virus by the Renilla luciferase assay system. GalT and ACBD3 FQ258AA were used as controls.