Structure of the Microtubule-Binding Domain of Flagellar Dynein

Kato, Yusuke; Yagi, Toshiki; Harris, Sarah A.; Ohki, Shiro; Yura, Kei; Shimizu, Yoshikazu; Honda, Shinya; Kamiya, Ritsu; Burgess, Simon; Tanokura, Masaru

doi:10.1016/j.str.2014.08.021

Cited by 21 publications

(34 citation statements)

References 43 publications

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“…Combining our current work with previous studies (Carter et al, 2008; Kato et al, 2014), we now have structures of two distantly related dynein MTBDs both on and off the microtubule. In both the cytoplasmic dynein-1 and DNAH7 MTBDs the predominant change is the position of H1, suggesting this is a universal mechanism for controlling dynein binding to microtubules.…”

Section: Discussionmentioning

confidence: 52%

“… C – A comparison between the microtubule bound DNAH7 model (violet) and the low-affinity Flagellar dynein C NMR structure ((Kato et al, 2014), 2RR7, ensemble chain 8, docked to the same density, white). As with cytoplasmic dynein, the only major conformational changes between the two is the upward movement of CC1/H1 (purple and grey for DNAH7 and 2RR7 respectively)…”

Section: Resultsmentioning

confidence: 99%

“…Following 3D classification and refinement, we obtained a 4.8Å resolution map ( Figure 3B, S4A, S7A ). A model of the DNAH7 MTBD was refined into the density using a homology model to the low-affinity NMR structure (PDB 2RR7, (Kato et al, 2014)) as a starting point ( Figure 3B, S7B, Table 2 ). Comparing the final microtubule-bound model to the original low-affinity structure, we observe the same conformational changes as for SRS-DYNC1H1 3260-3427 ( Figure 3C ).…”

Section: Resultsmentioning

confidence: 99%

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Cryo-EM of dynein microtubule-binding domains shows how an axonemal dynein distorts the microtubule

Lacey

Scheres

et al. 2019

Preprint

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Dyneins are motor proteins responsible for transport in the cytoplasm and the beating of the axoneme in cilia and flagella. They bind and release microtubules via a compact microtubule-binding domain (MTBD) at the end of a long coiled-coil stalk. Here we address how cytoplasmic and axonemal dynein MTBDs bind microtubules at near atomic resolution. We decorated microtubules with MTBDs of cytoplasmic dynein-1 and axonemal dynein DNAH7 and determined their cryo-EM structures using the stand-alone Relion package. We show the majority of the MTBD is remarkably rigid upon binding, with the transition to the high affinity state controlled by the movement of a single helix at the MTBD interface. In addition DNAH7 contains an 18-residue insertion, found in many axonemal dyneins, that reaches over and contacts the adjacent protofilament. Unexpectedly we observe that DNAH7, but not dynein-1, induces large distortions in the microtubule cross-sectional curvature. This raises the possibility that dynein coordination in axonemes is mediated via conformational changes in the microtubule.

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Section: Discussionmentioning

confidence: 52%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Cryo-EM of dynein microtubule-binding domains shows how an axonemal dynein distorts the microtubule

Lacey

Scheres

et al. 2019

Preprint

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“…We generated a construct encoding a 137-residue fragment of the yeast cytoplasmic dynein; the corresponding protein contains the entire microtubule-binding domain (MTBD) and the part of the coiled-coil ( Fig. 1a), as shown in a previous NMR study of mouse cytoplasmic dynein 22,26 . The protein (termed MTBD-WT) encoded by this construct was expressed in E. coli and purified to homogeneity.…”

Section: Construct Design Of Mtbds Stabilized In the Low-affinity Andmentioning

confidence: 99%

Structural basis for two-way communication between dynein and microtubules

Nishida

Komori

Takarada

et al. 2019

Preprint

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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 of dynein via an a-helical coiled-coil stalk, but the structural basis of this communication remains elusive. Here, we regulated MTBD either in high-affinity or low-affinity states by introducing a disulfide bond between the coiled-coils and analyzed the resulting structures by NMRand cryo-EM.In the MT-unbound state, the affinity changes of MTBD were achieved by sliding of the N-terminal α-helix by one half-turn, which suggests that structural changes propagate from the ATPase-domain to MTBD. In addition, cryo-EM analysis showed that MT binding induced further sliding of the N-terminal α-helix even without the disulfide bond, which suggests the MT-induced conformational changes propagate toward the ATPase domain. Based on differences in the MT-binding surface between the high-and low-affinity states, we propose a potential mechanism for the directional bias of dynein movement on MT tracks. Nishida et al.

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“…However, the atomic detail of the affinity switching, especially regarding how the helix sliding near the ATPase domain is transmitted to MTBD, remains elusive. Currently available crystal or NMR structures that include MTBD moiety [20][21][22] mostly represent the low-affinity conformation, even for the full-length motor domain of Dictyostelium discoideum cytoplasmic dynein in the ADP-bound state 9 , which would reflect the high-affinity state for MTs based on the biochemical observation. The cryo-EM-based atomic model of MT-bound MTBD in the high-affinity binding state 23,24 was used to infer that the conformational changes of the N-terminal H1 helix are essential for high-affinity binding.…”

mentioning

confidence: 99%

Structural basis for two-way communication between dynein and microtubules

et al. 2020

View full text Add to dashboard Cite

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.

show abstract

Structure of the Microtubule-Binding Domain of Flagellar Dynein

Cited by 21 publications

References 43 publications

Cryo-EM of dynein microtubule-binding domains shows how an axonemal dynein distorts the microtubule

Cryo-EM of dynein microtubule-binding domains shows how an axonemal dynein distorts the microtubule

Structural basis for two-way communication between dynein and microtubules

Structural basis for two-way communication between dynein and microtubules

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