The ATPase Pathway That Drives the Kinesin-14 Kar3Vik1 Powerstroke

Chen, Chun Ju; Porche, Ken; Rayment, Ivan; Gilbert, Susan P.

doi:10.1074/jbc.m112.395590

Cited by 18 publications

(28 citation statements)

References 51 publications

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“…S3; Table S1) (9,24) and was required for Ncd to detach from the MT (E6) (24). Interestingly, ATP hydrolysis for Kar3-Cik1 and Kar3Vik1 were also measured at 26 s −1 (27,28), reflecting the similarity of residues at the pivot point for the coiled-coil stalk rotation and the active site residue configuration for ATP hydrolysis. The presteady-state kinetics performed previously measured the dissociation step at 12-14 s −1 (Table S1) (23,24), and it is proposed that after detachment from the MT, the Ncd E6 intermediate returns to the E0 state, poised to rebind to the MT.…”

Section: Discussionmentioning

confidence: 99%

“…1, E0-E2). For Kar3Cik1 and Kar3Vik1, this strategy was necessary to capture the transient E1 state in which the MHD collided with the MT first (27,28). For these experiments, 10 μM Ncd•ADP was rapidly mixed in the stopped-flow instrument with 40 μM MTs plus the fluorescent ATP analog 2′-(3′)-O-(N-methylanthraniloyl) ATP (mantATP).…”

Section: Resultsmentioning

confidence: 99%

“…The pulse-chase experiments (Fig. S2) revealed a rapid ATP-promoted isomerization at 81 s −1 , which has been interpreted to represent the structural transitions required to form the intermediate poised for ATP hydrolysis (27,28). This postpowerstroke intermediate was captured by cryoEM using the nonhydrolyzable ATP analog, AMPPNP (10,12).…”

Section: Discussionmentioning

confidence: 99%

“…Through our recent studies on S. cerevisiae Kar3Vik1 and Kar3Cik1, we discovered novel properties of these yeast kinesin14s that challenged the earlier models of how kinesin-14s generate force for their cellular functions (13,15,(26)(27)(28). The C-terminal globular domain of Vik1 exhibits the structure of a kinesin motor domain (MD), yet Vik1 as well as Cik1 lack a nucleotide-binding site (13,26).…”

mentioning

confidence: 96%

“…S1). The following assumptions were made based on earlier studies (9,13,23,24,(26)(27)(28): (i) In solution, homodimeric Ncd exhibits an asymmetry in which one head holds ADP tightly bound while the partner head holds ADP weakly (E0) (24). (ii) MT collision is followed by rapid ADP release (E0-E1) (24).…”

mentioning

confidence: 99%

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Drosophila Ncd reveals an evolutionarily conserved powerstroke mechanism for homodimeric and heterodimeric kinesin-14s

Zhang¹,

Dai

Hahn

et al. 2015

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

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Drosophila melanogaster kinesin-14 Ncd cross-links parallel microtubules at the spindle poles and antiparallel microtubules within the spindle midzone to play roles in bipolar spindle assembly and proper chromosome distribution. As observed for Saccharomyces cerevisiae kinesin-14 Kar3Vik1 and Kar3Cik1, Ncd binds adjacent microtubule protofilaments in a novel microtubule binding configuration and uses an ATP-promoted powerstroke mechanism. The hypothesis tested here is that Kar3Vik1 and Kar3Cik1, as well as Ncd, use a common ATPase mechanism for force generation even though the microtubule interactions for both Ncd heads are modulated by nucleotide state. The presteady-state kinetics and computational modeling establish an ATPase mechanism for a powerstroke model of Ncd that is very similar to those determined for Kar3Vik1 and Kar3-Cik1, although these heterodimers have one Kar3 catalytic motor domain and a Vik1/Cik1 partner motor homology domain whose interactions with microtubules are not modulated by nucleotide state but by strain. The results indicate that both Ncd motor heads bind the microtubule lattice; two ATP binding and hydrolysis events are required for each powerstroke; and a slow step occurs after microtubule collision and before the ATP-promoted powerstroke. Note that unlike conventional myosin-II or other processive molecular motors, Ncd requires two ATP turnovers rather than one for a single powerstroke-driven displacement or step. These results are significant because all metazoan kinesin-14s are homodimers, and the results presented show that despite their structural and functional differences, the heterodimeric and homodimeric kinesin-14s share a common evolutionary structural and mechanochemical mechanism for force generation.presteady-state kinetics | dynamic modeling | microtubules I n the early stages of mitosis and meiosis, the bipolar metaphase spindle must be established, and kinesin-14 molecular motors play key roles in this process (1-4). In contrast to the microtubule (MT) plus-end directed processive kinesins, kinesin-14s are not processive as single molecules; they promote MT minus-enddirected force and use an ATP-promoted powerstroke to crosslink and slide one MT relative to another (5-17). Sequence analysis indicates that all members of the kinesin-14 subfamily are dimeric, yet the structural organization of kinesin-14 motors differs from the N-terminal processive kinesins. The kinesin-14s exhibit C-terminal motor domains connected by an N-terminal continuous coiled-coil stalk with an N-terminal ATP-independent MT binding site (7,(18)(19)(20)(21)(22). And although most of the kinesin-14s are homodimeric, some yeast species, including Saccharomyces cerevisiae and Candida glabrata, contain heterodimeric kinesin-14s (13,14,19). The conventional hypothesis for homodimeric kinesin-14 force generation proposed that only one motor head interacts with the MT and only one ATP turnover is required to complete the powerstroke (Fig. S1A) (10, 12). In contrast, our presteady-state kinetics (9,...

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Section: Discussionmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 96%

mentioning

confidence: 99%

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Drosophila Ncd reveals an evolutionarily conserved powerstroke mechanism for homodimeric and heterodimeric kinesin-14s

Zhang¹,

Dai

Hahn

et al. 2015

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

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Kinesin Motor Enzymology: Chemistry, Structure, and Physics of Nanoscale Molecular Machines

Cochran

2015

Biophys Rev

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Molecular motors are enzymes that convert chemical potential energy into controlled kinetic energy for mechanical work inside cells. Understanding the biophysics of these motors is essential for appreciating life as well as apprehending diseases that arise from motor malfunction. This review focuses on kinesin motor enzymology with special emphasis on the literature that reports the chemistry, structure and physics of several different kinesin superfamily members.

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Kinesin-14 KIFC1 promotes acrosome formation and chromatin maturation during mouse spermiogenesis

Wei

Fan

Lin

et al. 2023

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research

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The ATPase Pathway That Drives the Kinesin-14 Kar3Vik1 Powerstroke

Cited by 18 publications

References 51 publications

Drosophila Ncd reveals an evolutionarily conserved powerstroke mechanism for homodimeric and heterodimeric kinesin-14s

Drosophila Ncd reveals an evolutionarily conserved powerstroke mechanism for homodimeric and heterodimeric kinesin-14s

Kinesin Motor Enzymology: Chemistry, Structure, and Physics of Nanoscale Molecular Machines

Kinesin-14 KIFC1 promotes acrosome formation and chromatin maturation during mouse spermiogenesis

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