Extended Data Fig. 5: Additional analysis of human MT-U and MT-B cryo-EM structures.

(a) Stick representations of the dynein_MOTOR^MT-B AAA sites showing the nucleotide electron density (the center of each image) and surrounding residues (residues are color-coded as shown in panel b schematic). (b) Vector maps depicting pairwise alpha carbon interatomic distances between the dynein_MOTOR^MT-B with the following: apo yeast dynein (4AKG), AMPPNP-bound yeast dynein (4W8F), ADP-bound Dictyostelium discoideum dynein (3VKG), and ADP-Vi and Pac1-bound yeast dynein (7MGM)^37,40,79,81. Structures were globally aligned after removal of the linkers. (c) AAA1-AAA2L domains from dynein_MOTOR^MT-B (shades of green) and the native microtubule-bound dynein-1 (7Z8G; magenta and pink) overlaid to depict the high degree of structural similarity. The two were locally aligned using AAA1. (d) AAA3-AAA4L domains from dynein_MOTOR^MT-B (shades of green) overlaid with either the native microtubule-bound dynein-1 (left, magenta and pink) or the ADP-bound Dictyostelium discoideum dynein (right, blue and green). (e and f) Stick representations of the LIS1-unbound (e) or bound (f) dynein_MOTOR^MT-U AAA sites showing the nucleotide electron density and surrounding residues (residues are color-coded as shown in panel b schematic). (g) Vector maps depicting pairwise alpha carbon interatomic distances between the LIS1-bound dynein_MOTOR^MT-U with those described for panel b. Structures were globally aligned after removal of the linkers. (h) AAA1-AAA2L domains from dynein_MOTOR^MT-U (left) and dynein_MOTOR^MT-B (right) with residues of dynein_MOTOR^MT-U contacting the Vi highlighted (E1959, Walker B; N2019, Sensor-I; R2358, arginine finger; N2316; A2354;). Distances between these residues and the Vi (or, for the dynein_MOTOR^MT-B structure, between them and where the Vi would be) are indicated.