Fig. 6: Lis1 binding to dynein is blocked by the Lis1-α2 interaction in the absence of Nde1. | Nature Communications

Fig. 6: Lis1 binding to dynein is blocked by the Lis1-α2 interaction in the absence of Nde1.

From: Nde1 promotes Lis1-mediated activation of dynein

Fig. 6

a Structures of Lis1-DHC (PDB ID: 8DYV63), Lis1-α2 (PDB ID: 1VYH14), and Lis1-Nde11–190 (predicted by AlphaFold2). Lis1 residues R316 and W340 that facilitate binding to dynein are also critical for binding to Nde1 and α2. b Mass photometry reveals that α2 forms a complex with Lis1, but not with mtLis1, and does not simultaneously bind to Lis1 and Nde11–190. A multiple Gaussian fit predicts the average mass and percentage of each population (solid curves, Supplementary Table 2). c Normalized absorbance (top) and fluorescence picture of a denaturing gel (bottom) of the mixtures of Lis1-LD655, Nde11–190-LD555, and α-Alexa488 eluted from a size exclusion column. d (Left) Ratios of surface-immobilized dynein colocalized with 1 nM Lis1 under increasing concentrations of α2. (Right) Ratios of surface-immobilized Lis1 colocalized with 25 nM Nde1 under increasing concentrations of α2. From left to right, n = 10, 10, 10, 14, and 19 (Left) and 9, 7, 7, 6, and 6 (Right) imaging areas (40 µm × 40 µm) with minimum 100 dyneins (Left) and Lis1 (Right). e Ratios of surface-immobilized Lis1 colocalized with 10 nM α2 under increasing concentrations of Nde11–190. For each condition, n = 12 imaging areas (40 µm × 40 µm) with minimum 40 Lis1. f Kymographs show wtDDR motility under 0–50 nM α2 and 0–10 nM Lis1 without Nde1. g The run frequency of wtDDR under 0–50 nM α2 and 0–100 nM Lis1 without Nde1. Results were normalized to no Lis1 and α2 condition. From left to right, n = 22, 20, 20, and 20 microtubules. h Kymographs show wtDDR motility under different concentrations of α2 and Nde1 in 10 nM Lis1. i The run frequency of wtDDR under different concentrations of α2 and Nde1 in 10 nM Lis1 (mean ± s.d.; n = 20 microtubules for each condition). Results were normalized to the 10 nM Lis1 condition. In (d, e, g), the center line and whiskers represent the mean and s.d., respectively. P values are calculated from a two-tailed t test. Source data are provided as a Source Data file.

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