Structural basis for two-way communication between dynein and microtubules

Abstract

The movements of cytoplasmic dynein on microtubule (MT) tracks is achieved by two-way communication between the microtubule-binding domain (MTBD) and the ATPase domain via a coiled-coil stalk, but the structural basis of this communication remains elusive. Here, we regulate MTBD either in high-affinity or low-affinity states by introducing a disulfide bond to the stalk and analyze the resulting structures by NMR and cryo-EM. In the MT-unbound state, the affinity changes of MTBD are achieved by sliding of the stalk α-helix by a half-turn, which suggests that structural changes propagate from the ATPase-domain to MTBD. In addition, MT binding induces further sliding of the stalk α-helix even without the disulfide bond, suggesting how the MT-induced conformational changes propagate toward the ATPase domain. Based on differences in the MT-binding surface between the high-and lowaffinity states, we propose a potential mechanism for the directional bias of dynein movement on MT tracks.

Structures of outer-arm dynein array on microtubule doublet reveal a motor coordination mechanism

“…First, an 18-degree rotation between the β-stalk and β-MTBD is required for microtubule-binding state alteration (Fig. 5a and 5b), whereas β-MTBD remains in the high microtubule-affinity s tate (42) (Fig. 5a and 5c).…”

Three-color single-molecule imaging reveals conformational dynamics of dynein undergoing motility

“…ssDNA, single-stranded DNA. ( B ) Structure of a yeast cytoplasmic dynein MTBD (gold) bound to tubulin (gray) [Protein Data Bank ID code 6KIQ ( 56 )]. The exact position of the YBBR-tag insertion is shown as red spheres.…”

Regulatory mechanisms of the dynein-2 motility by post-translational modification revealed by MD simulation

“…This model was already contained whole motor domain, and the sequence was DYHC2, so we used MODELLER only for modeling disordered region. Also, for the ADP state dynein which took high-affinity MTBD was modeled by combining the three models taken from the PDB ID: 3VKH (44) for AAA+ and stalk, 3J1T (45) for stalk and MTBD, and 6KIQ (46) for MTBD and the interface with MT in the following protocol. At first, 3VKH was the X-ray crystal structure of Dictyostelium discoideum cytoplasmic dynein-1.…”