Fig. 6: The NTF2 domain is required to target Bd1075 to the convex cell face and generate curvature.
From: Asymmetric peptidoglycan editing generates cell curvature in Bdellovibrio predatory bacteria
a B. bacteriovorus Bd1075-mCherry double-crossover (DXO) attack-phase cells (left), showing the localization of wild-type Bd1075-mCherry to the convex cell face, and representative of 3 biological repeats. Dashed boxed region is shown in a close-up (middle). Scale bars = 2 μm. Heatmap (right) depicts the location of wild-type Bd1075-mCherry foci detected in n = 1189 cells from 3 biological repeats. White-yellow = highest intensity, purple-black = lowest intensity. b Schematics of Bd1075-mCherry single-crossover constructs used in c–e. Full-length: Residues 1–329 (wild-type complete protein) fused to mCherry, A304: Residues 1–304 (contains a completed NTF2 domain including the B. bacteriovorus-specific residue W303) fused to mCherry, E302: Residues 1–302 (does not complete the NTF2 domain) fused to mCherry, C156A: Residues 1–329 (full-length with a point mutation of C156A in the catalytic LD-CPase domain) fused to mCherry, and Y274A: Residues 1–329 (full-length with a point mutation of Y274A in the NTF2 domain) fused to mCherry. c, d Bd1075-mCherry single-crossover constructs introduced into either B. bacteriovorus HD100 c wild-type (contains a native wild-type copy of bd1075) or d Δbd1075 (lacking a wild-type copy of bd1075). Attack-phase cell images and adjacent heatmaps show targeting of Bd1075-mCherry. Images and heatmaps were generated from 3 biological repeats (n = number of cells analyzed). Scale bars = 2 μm. e Curvature measurements of B. bacteriovorus Δbd1075 attack-phase cells containing different single-crossover Bd1075-mCherry fusions. n = 2099 cells (WT HD100), 1886 cells (Δbd1075), 2577 cells (Δbd1075 (FullmCh)), 2170 cells (Δbd1075 (A304mCh)), 2812 cells (Δbd1075 (E302mCh)), 2083 cells (Δbd1075 (C156AmCh)), or 2523 cells (Δbd1075 (Y274AmCh)) per strain from 3 biological repeats. Error bars represent 95% confidence intervals of the median. All pairwise comparisons between strains (except for Δbd1075 vs Δbd1075 (C156AmCh) were significant (p < 0.0001; Kruskal–Wallis test with Dunn’s multiple comparisons). Frequency distributions are included in Supplementary Fig. 5c. Source Data are provided as a Source Data file.
