Fig. 4: GlpR links the mid-cell divisome with the future polar elongasome via Wag31.
a, Left: representative images of GlpR-mNeon expressed in Cglu and Cglu_Δglpr. Right: heat map of the localization of GlpR-mNeon in Cglu (n = 111). b, Violin plots (cell length). Cohen’s d, from top to bottom: (**d = 0.98, P = 2.88 × 10−85), (nsd = 0.49, P = 6.34 × 10−19), (nsd = 0.39, P = 2.78 × 10−13). The expression levels of GlpR-mNeon are shown in Extended Data Fig. 4d. c, Representative images of Cglu_Δglpr complementation (overexpression conditions). d, Violin plots (cell surface areas). Cohen’s d: (****d = 3.95, P = 7.34 × 10−63), (****d = 2.35, P = 1.01 × 10−46), (***d = 1.4, P = 5.37 × 10−319). The expression levels of GlpR-mNeon are shown in Extended Data Fig. 4e. e, Violin plots of cell length. Cohen’s d: (**d = 1.17, P = 7.22 × 10−255), (*d = 1.70, P = 6.20 × 10−138), (*d = 0.61, P = 3.07 × 10−60), (nsd = 0.49, P = 8.12 × 10−34). Boxes indicate the 25th to the 75th percentile, mean and median indicated with a dot and a line, respectively, in the box. Number of cells used (n) below the violin representation corresponds to triplicates. f, Histogram of number of septa per cell, calculated from n cells from 3 independent experiments (Cglu_Δglp, n = 873, 1,538 and 840; Cglu, n = 718, 1,468 and 1,223; Cglu_Δglp + GlpRΔIDR2, n = 451, 737 and 841); open circles represent the corresponding data points; mean ± s.d.; Cohen’s d, from top to bottom: (***d = 1.60, P = 0), (**d = 0.80, P = 3.70 × 10−169), (**d = 0.79, P = 8.20 × 10−147). g, Co-IP of GlpR-Wag31 for indicated strains using GlpR as bait. Total (T), wash (W) and elution (E) fractions were analysed by western blot using α-GlpR and α-Wag31 antibodies. Arrows indicate GlpR (top) and Wag31 (bottom) and Wag31 is additionally highlighted by a red *. The black bar corresponds to the 55 kDa molecular weight marker. h, Wag31 recovery in co-IPs of Glp from indicated strains. Each point corresponds to the normalized XIC intensity in each biologically independent replicate (n = 4) for each condition; mean ± s.d. Wag31 FC = 1.78 (P = 0.004). Statistical analysis was performed using a two-sided unpaired Student’s t-test. i, BLI sensorgrams of Wag31 binding to GlpR. Wag31 concentrations: 150 μM (dark blue) to 2.3 μM (light green) in 2-fold dilutions.
