A flipped ion pair at the dynein–microtubule interface is critical for dynein motility and ATPase activation

Uchimura, Seiichi; Fujii, Takashi; Takazaki, Hiroko; Ayukawa, Rie; Nishikawa, Yosuke; Minoura, Itsushi; Hachikubo, You; Kurisu, Genji; Sutoh, Kazuo; Kon, Takahide; Namba, Keiichi; Muto, Etsuko

doi:10.1083/jcb.201407039

Cited by 45 publications

(75 citation statements)

References 55 publications

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“…Experiments by Gibbons et al [31] first suggested that the “α-registry” is associated with a strong binding state of the MTBD and a four residue shifted “β+-registry” is associated with a weak binding state. Several subsequent experiments have provided additional support for this hypothesis [29, 32–35]. The sliding of one of the helices of the stalk between the α- and β-registries is thought to provide a mechanism for communicating between the AAA ring and the MTBD.…”

Section: The Dynein Motor Domain and Its Conformational Changesmentioning

confidence: 98%

“…Similar to this model, dynein has been shown to undergo one-dimensional diffusion along the microtubule surface in its weakly bound (ATP) state. This result suggests that the MTBD can hold on to the microtubule weakly and that thermal energy can displace the MTBD from one tubulin subunit to the next along the microtubule [35, 53, 54]. Furthermore, optical trap studies have uncovered a directional asymmetry in the binding of dynein to the microtubules.…”

Section: How Dynein Steps Towards the Microtubule Minus Endmentioning

confidence: 99%

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How Dynein Moves Along Microtubules

Bhabha

Johnson

Schroeder

et al. 2016

Trends in Biochemical Sciences

130

127

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Cytoplasmic dynein, a member of the AAA family of ATPases, drives the processive movement of numerous intracellular cargos towards the minus end of microtubules. Here, we summarize the structural and motile properties of dynein and highlight features that distinguish this motor from kinesin-1 and myosin V, two well-studied transport motors. Integrating information from recent crystal and cryo-EM structures as well as high-resolution single molecule studies, we also discuss models for how dynein biases its movement in one direction along a microtubule track, and present a movie that illustrates these principles.

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Section: The Dynein Motor Domain and Its Conformational Changesmentioning

confidence: 98%

Section: How Dynein Steps Towards the Microtubule Minus Endmentioning

confidence: 99%

How Dynein Moves Along Microtubules

Bhabha

Johnson

Schroeder

et al. 2016

Trends in Biochemical Sciences

130

127

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“…The assumption that isoforms or post‐translational states can be averaged or overlooked appears to be untrue. Recent work has shown that sequence and modification state can change the dynamics, and the binding of associated proteins and enzymes . Recombinant tubulin from yeast can achieve high purity and has been systematically studied with various mutants .…”

Section: Gtpase Filaments: Microtubules and Tubulin Structure And Funmentioning

confidence: 99%

Invited review: Microtubule severing enzymes couple atpase activity with tubulin GTPase spring loading

Bailey¹,

Jiang

Dima

et al. 2016

Biopolymers

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Microtubules are amazing filaments made of GTPase enzymes that store energy used for their own self-destruction to cause a stochastically driven dynamics called dynamic instability. Dynamic instability can be reproduced in vitro with purified tubulin, but the dynamics do not mimic that observed in cells. This is because stabilizers and destabilizers act to alter microtubule dynamics. One interesting and understudied class of destabilizers consists of the microtubule-severing enzymes from the ATPases Associated with various cellular Activities (AAA+) family of ATP-enzymes. Here we review current knowledge about GTP-driven microtubule dynamics and how that couples to ATP-driven destabilization by severing enzymes. We present a list of challenges regarding the mechanism of severing, which require development of experimental and modeling approaches to shed light as to how severing enzymes can act to regulate microtubule dynamics in cells. © 2016 Wiley Periodicals, Inc. Biopolymers 105: 547-556, 2016.

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“…To investigate whether there are specific residues in tubulin that are shared in the interaction with kinesin, PRC1, and dynein, we performed sequence and structural alignments for these proteins, using tubulin as the common element to superimpose the MT-MAP models (Materials and Methods). Although as yet there is no atomic-resolution information on the binding of the dynein MTBD to MTs, hybrid models of this interaction have been generated by integrating diverse sources of data, including crystal structures (41-47), medium-resolution EM maps (43,47), and biochemical data (48,49). Superposition of the MT-kinesin atomic model with our MT-PRC1-SC atomic model identified kinesin R278 and PRC1 R381 as functionally similar in their interaction with tubulin (potentially with β-tubulin D420 and β-tubulin E427) (Fig.…”

Section: Prc1's Disordered C-terminal Domain Forms Electrostatic Intementioning

confidence: 99%

“…Previous atomic models of motor domains bound to MTs derived from cryo-EM reconstructions (43,(45)(46)(47) and crystallography (41) were superimposed on our final atomic model using the H11′ helix on α-tubulin to align the binding pockets locally. The following structures were inspected to identify tubulin residues that potentially share binding activity for PRC1, kinesin, or dynein: 2P4N (42), 4HNA (41), 4CK5 (44), 4UXO (45), 3J8Y (46), 3J6P (47), 3J1U, and 3J1T (43).…”

Section: Simulationsmentioning

confidence: 99%

Near-atomic cryo-EM structure of PRC1 bound to the microtubule

Kellogg

Howes

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Proteins that associate with microtubules (MTs) are crucial to generate MT arrays and establish different cellular architectures. One example is PRC1 (protein regulator of cytokinesis 1), which cross-links antiparallel MTs and is essential for the completion of mitosis and cytokinesis. Here we describe a 4-Å-resolution cryo-EM structure of monomeric PRC1 bound to MTs. Residues in the spectrin domain of PRC1 contacting the MT are highly conserved and interact with the same pocket recognized by kinesin. We additionally found that PRC1 promotes MT assembly even in the presence of the MT stabilizer taxol. Interestingly, the angle of the spectrin domain on the MT surface corresponds to the previously observed cross-bridge angle between MTs cross-linked by full-length, dimeric PRC1. This finding, together with molecular dynamic simulations describing the intrinsic flexibility of PRC1, suggests that the MT-spectrin domain interface determines the geometry of the MT arrays cross-linked by PRC1.ells rely on the microtubule (MT) cytoskeleton to help organize organelles (1), control cellular morphology (2), provide mechanical stability (3, 4), and form the spindle apparatus used to segregate chromosomes during cell division (5-8). The diverse functions of MTs are made possible through the action of motors (i.e., kinesin and dynein) and nonmotor MT-associated proteins (MAPs) that tightly regulate the MT network. Some of these proteins act by binding directly to the ends of MTs, either to the highly dynamic plus end, such as the conserved end-binding proteins (9, 10), or to the minus end (11,12). Other MT regulators, such as MAP1 and MAP2/Tau, bind along the MT lattice, stabilizing it and helping build parallel arrays, most notably in axons (13,14). Members of the MAP65 family, which includes human protein regular of cytokinesis 1 (PRC1) and its budding yeast ortholog Ase1, form antiparallel MT arrays important for setting the spindle midzone and determining the location of the cytokinetic ring (8,(15)(16)(17)(18)(19). In addition to binding selectively to antiparallel MTs, PRC1 recruits other spindle-organizing factors and therefore is an essential component of the mitotic spindle (15,20). Proper functioning of PRC1 requires cell-cycle-dependent localization and regulation. PRC1 contains two nuclear localization signals (NLSs) in its C-terminal domain (Fig. 1A) and is found almost exclusively in the nucleus during interphase (21-23). As the cell enters mitosis, PRC1 localizes to the spindle and becomes concentrated at the midzone by late anaphase (22), similar to observations for Ase1 (24). PRC1 is subject to phosphorylation by several cyclin-cyclin-dependent kinase (CDK) complexes (21) and together with kinesin-4 can regulate MT antiparallel overlap at the spindle midzone (20,23,25,26).Previous studies have identified that the dimerization domain of PRC1 is within the N-terminal domain (Fig. 1A) and that the primary MT binding region relies on a set of conserved residues within the central spectrin domain (PRC1-S) (15,22,2...

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A flipped ion pair at the dynein–microtubule interface is critical for dynein motility and ATPase activation

Abstract: Salt bridges at the dynein–microtubule interface couple microtubule binding to ATPase activation and thereby control the directional movement of dynein

Cited by 45 publications

References 55 publications

How Dynein Moves Along Microtubules

How Dynein Moves Along Microtubules

Invited review: Microtubule severing enzymes couple atpase activity with tubulin GTPase spring loading

Near-atomic cryo-EM structure of PRC1 bound to the microtubule

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