Directionally biased sidestepping of Kip3/kinesin-8 is regulated by ATP waiting time and motor–microtubule interaction strength

Mitra, Aniruddha; Ruhnow, Felix; Girardo, Salvatore; Diez, Stefan

doi:10.1073/pnas.1801820115

Cited by 47 publications

(76 citation statements)

References 67 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…(Note that after ATP binding the NL docking makes the ADP‐head move forward with the high probability, resulting in forward stepping.) Moreover, it is expected that the longer the lifetime in 1HB state before ATP binding, the larger the sidestepping probability, which is also consistent with the recent experimental data …”

Section: Discussionsupporting

confidence: 90%

“…Our simulation results showing that in 1HB state the detached ADP‐head is in the leftward position relative to the MT‐bound nucleotide‐free head can also explain well the available experimental data showing that kinesin dimers sidestep with a directional bias toward the left . In 1HB state before ATP binding to the MT‐bound head, the ADP‐head can detach from the MT‐bound head by overcoming the high affinity between the two heads and then occasionally bind to MT, releasing ADP.…”

Section: Discussionsupporting

confidence: 81%

“…Moreover, it is expected that the longer the lifetime in 1HB state before ATP binding, the larger the sidestepping probability, which is also consistent with the recent experimental data. 67…”

Section: T a B L E 2 Statistical Results Of Salt-bridge Interaction Bmentioning

confidence: 99%

See 2 more Smart Citations

All‐atom molecular dynamics simulations reveal how kinesin transits from one‐head‐bound to two‐heads‐bound state

et al. 2019

View full text Add to dashboard Cite

Kinesin dimer walks processively along a microtubule (MT) protofilament in a hand‐over‐hand manner, transiting alternately between one‐head‐bound (1HB) and two‐heads‐bound (2HB) states. In 1HB state, one head bound by adenosine diphosphate (ADP) is detached from MT and the other head is bound to MT. Here, using all‐atom molecular dynamics simulations we determined the position and orientation of the detached ADP‐head relative to the MT‐bound head in 1HB state. We showed that in 1HB state when the MT‐bound head is in ADP or nucleotide‐free state, with its neck linker being undocked, the detached ADP‐head and the MT‐bound head have the high binding energy, and after adenosine triphosphate (ATP) binds to the MT‐bound head, with its neck linker being docked, the binding energy between the two heads is reduced greatly. These results reveal how the kinesin dimer retains 1HB state before ATP binding and how the dimer transits from 1HB to 2HB state after ATP binding. Key residues involved in the head‐head interaction in 1HB state were identified.

show abstract

Section: Discussionsupporting

confidence: 90%

Section: Discussionsupporting

confidence: 81%

See 1 more Smart Citation

All‐atom molecular dynamics simulations reveal how kinesin transits from one‐head‐bound to two‐heads‐bound state

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Interestingly several mitotic kinesins, like kinesin-5 5 , kinesin-8 6,7 and kinesin-14 8,9 , have been shown to exhibit off-axis components in their stepping behavior, i.e. they do not move strictly parallel to the longitudinal microtubule axis.…”

Section: Introductionmentioning

confidence: 99%

“…Ncd is a non-processive, minus-end directed motor possessing an additional diffusive microtubule-interacting site in its N-terminal tail domain that enables the motor to crosslink and slide anti-parallel microtubules 10 . Specifically, we suspend long microtubules on nano-fabricated ridges 7,13 and track the 3D motion of short microtubules along the long ones. We discover that anti-parallel microtubules do both, rotate around their own axes and helically move around each other, in a right-handed manner.…”

Section: Introductionmentioning

confidence: 99%

The kinesin-14, Ncd, drives a right-handed, helical motion of antiparallel microtubules around each other

Mitra

Gandhimathi

Ruhnow

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

Within the mitotic spindle, several kinesin motors crosslink and slide microtubules. While some of them (e.g. kinesin-5, kinesin-8 and kinesin-14) have been shown to exhibit sideways components in their step cycles, the impact of the resulting off-axis power strokes on motility and force generation in the spindle has not been investigated so far. Here, we develop and utilize a novel three-dimensional in vitro motility assay to explore the kinesin-14, Ncd, driven sliding of crosslinked, fluorescently-labeled microtubules. We find that free microtubules, sliding in an antiparallel orientation on microtubules suspended between nanofabricated ridges, not only rotate around their own axis but also move around the suspended microtubules with right-handed helical trajectories. In contrast, microtubules crosslinked in parallel orientation are static with neither longitudinal nor helical motion. Further, our technique allows us to measure the in situ spatial extension of the motors between the crosslinked microtubules to be about 20 nm. We argue that the capability of microtubule-crosslinking kinesins to cause helical motion of microtubules around each other allows for flexible filament organization, roadblock circumvention and torque generation in the mitotic spindle.

show abstract

N‐terminal β‐strand of single‐headed kinesin‐1 can modulate the off‐axis force‐generation and resultant rotation pitch

et al. 2020

View full text Add to dashboard Cite

In in vitro microtubule gliding assays, most kinesins drive the rotation of gliding microtubules around their longitudinal axes in a corkscrew motion. The corkscrewing pitch is smaller than the supertwisted protofilament pitch of microtubules, indicating that the corkscrewing pitch is an inherent property of kinesins. To elucidate the molecular mechanisms through which kinesins corkscrew the microtubule, we performed three-dimensional tracking of a quantum dot bound to a microtubule translocating over a surface coated with single-headed kinesin-1 s under various assay conditions to alter the interactions between the kinesin and microtubule. Although alternations in kinesin concentration, ionic strength, and ATP concentration changed both gliding and rotational velocities, the corkscrewing pitch remained left-handed and constant at $0.3 μm under all tested conditions apart from a slight increase in pitch at a low ATP concentration. We then used our system to analyze the effect of point mutations in the N-terminal β-strand protruding from the kinesin motor core and found mutations that decreased the corkscrewing pitch. Our findings confirmed that the corkscrewing motion of microtubules is caused by the intrinsic properties of the kinesin and demonstrates that changes in the active or retarding force originating from the N-terminal β-strand in the head modulate the pitch.

show abstract

Directionally biased sidestepping of Kip3/kinesin-8 is regulated by ATP waiting time and motor–microtubule interaction strength

Cited by 47 publications

References 67 publications

All‐atom molecular dynamics simulations reveal how kinesin transits from one‐head‐bound to two‐heads‐bound state

All‐atom molecular dynamics simulations reveal how kinesin transits from one‐head‐bound to two‐heads‐bound state

The kinesin-14, Ncd, drives a right-handed, helical motion of antiparallel microtubules around each other

N‐terminal β‐strand of single‐headed kinesin‐1 can modulate the off‐axis force‐generation and resultant rotation pitch

Contact Info

Product

Resources

About