Fig. 5. Role of Chi in human dynein’s ability to move on microtubules.

a, A model of the human Chi AAA6₂ and AAA5₂–LIS1₁ interface. The arrow in the inset indicates the area highlighted in the main panel. b, Single-molecule velocity (median ± interquartile range) of TMR–dynein–dynactin–BICD2 complexes in the absence (white circles) or presence (black circles) of different human LIS1 constructs. The data points are represented as triangles, circles, squares, and hexagons corresponding to single measurements within each technical replicate. Four technical replicates were collected for each condition, and the number of data points (n) per each replicate is listed (no LIS1, n = 142, 91, 152, 131; LIS1, n = 147, 118, 119, 120; LIS1^{N203A D205A Y225A}, n = 149, 91, 120, 103; LIS1^{N203A D205A D245A}, n = 147 109 128 108). No LIS1 and LIS1 ***P = 0.0002, LIS1 and LIS1^{N203A D205A Y225A} **P = 0.0051, LIS1 and LIS1^{N203A D205A D245A} **P = 0.004. One-Way ANOVA with Tukey’s multiple comparison test. c, Processive runs (mean ± s.e.m.) of TMR–dynein–dynactin–BICD2 complexes in the absence (white circle) or presence (black circle) of different unlabeled human LIS1 constructs. The data points are represented as triangles, circles, squares, and hexagons corresponding to single measurements within each technical replicate. Four replicates per condition were collected, and the number of data points (n) per each replicate is listed (no LIS1, n = 14, 20, 20, 8; LIS1, n = 7, 16, 7, 5; Lis1^{N203A D205A Y225A}, n = 15, 7, 16, 9; Lis1^{N203A, D205A D245A}, n = 17, 9, 17, 9). No LIS1 and LIS1 *P = 0.023. One-Way ANOVA with Tukey’s multiple comparison test. d, Representative kymographs from single-molecule motility assays with purified TMR–dynein–dynactin–BICD2 in the absence (white circle) or presence (black circle) of different human LIS1 constructs. Scale bars, 10 μm (x) and 40 s (y). e, Schematic of the hypothesis for how Lis1 relieves dynein autoinhibition.

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