Fig. 2: Kelch13 mediates the protein interaction network at the micropore.
a Conservation of micropore proteins in representative species of the apicomplexans. The protein names with the corresponding ID (in brackets) were listed, followed by CRISPR fitness scores and the heatmap55 by the side of a conservation heatmap. The heatmaps were correspondingly scaled by –log E values and CRISPR scores. b Peptide numbers of IMC proteins and microtubules identified by proximity biotin labeling. c The proximity-based interactome of micropore proteins, showing interactions of the bait (deep blue) and the prey (light blue) filtered through a R-based SFINX package for 7 TurboID fusions (N = 2 datasets for each) with strictness = 4 (p value < 0.00005). A hypergeometric test was used in the SFINX analysis. The core nodes K13, PPG1 and EPS15 shared the same set of interactors. d–f Plaque formation of AID lines grown in ±IAA for 7 days. EPS15, PPG1 and K13 were separately fused with AID in tKD. Plaque numbers and sizes were scored and plotted (e, f). The sizes were measured for 18 plaques in each independent experiment. Scale = 0.5 cm. g Parasite replication for ±IAA parasites for 24 h, showing ratios of different parasites/vacuole in the total (n > 100 for each replicate). In comparing EPS15-AID +IAA vs. TIR1 −IAA, p < 0.0001 for 4 and 8 parasites/vacuole and p < 0.01 for 2 parasites/vacuole; in comparing AID-K13+ IAA vs. TIR1 −IAA and tKD +IAA vs. TIR1 −IAA, p < 0.0001 for 2, 4 and 8 parasites/vacuole. Auxin treatment removed all the 16 parasites/vacuole for the AID lines. e–g Mean ± SD for three independent experiments with triplicates, analyzed by one-way or two-way ANOVA with Tukey’s multiple comparison, ****p < 0.0001.
