A Homotetrameric Kinesin-5, KLP61F, Bundles Microtubules and Antagonizes Ncd in Motility Assays

Li Tao, Alex Mogilner, Gul Civelekoglu-Scholey, Roy Wollman, James Evans, Henning Stahlberg, Jonathan M. Scholey

Research output: Contribution to journalArticle

Abstract

Background: Mitosis depends upon the cooperative action of multiple microtubule (MT)-based motors. Among these, a kinesin-5, KLP61F, and the kinesin-14, Ncd, are proposed to generate antagonistic-sliding forces that control the spacing of the spindle poles. We tested whether purified KLP61F homotetramers and Ncd homodimers can generate a force balance capable of maintaining a constant spindle length in Drosophila embryos. Results: Using fluorescence microscopy and cryo-EM, we observed that purified full-length, motorless, and tailless KLP61F tetramers (containing a tetramerization domain) and Ncd dimers can all cross-link MTs into bundles in MgATP. In multiple-motor motility assays, KLP61F and Ncd drive plus-end and minus-end MT sliding at 0.04 and 0.1 μm/s, respectively, but the motility of either motor is decreased by increasing the mole fraction of the other. At the "balance point," the mean velocity was zero and MTs paused briefly and then oscillated, taking ∼0.3 μm excursions at ∼0.02 μm/s toward the MT plus end and then the minus end. Conclusions: The results, combined with quantitative analysis, suggest that these motors could act as mutual brakes to modulate the rate of pole-pole separation and could maintain a prometaphase spindle displaying small fluctuations in its steady-state length.

Original languageEnglish (US)
Pages (from-to)2293-2302
Number of pages10
JournalCurrent Biology
Volume16
Issue number23
DOIs
StatePublished - Dec 5 2006

Fingerprint

Kinesin
kinesin
Microtubules
microtubules
Assays
Poles
assays
Prometaphase
Spindle Poles
Fluorescence Microscopy
Mitosis
Drosophila
Fluorescence microscopy
Force control
Embryonic Structures
Adenosine Triphosphate
fluorescence microscopy
Brakes
Dimers
mitosis

Keywords

  • CELLBIO

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)

Cite this

Tao, L., Mogilner, A., Civelekoglu-Scholey, G., Wollman, R., Evans, J., Stahlberg, H., & Scholey, J. M. (2006). A Homotetrameric Kinesin-5, KLP61F, Bundles Microtubules and Antagonizes Ncd in Motility Assays. Current Biology, 16(23), 2293-2302. https://doi.org/10.1016/j.cub.2006.09.064

A Homotetrameric Kinesin-5, KLP61F, Bundles Microtubules and Antagonizes Ncd in Motility Assays. / Tao, Li; Mogilner, Alex; Civelekoglu-Scholey, Gul; Wollman, Roy; Evans, James; Stahlberg, Henning; Scholey, Jonathan M.

In: Current Biology, Vol. 16, No. 23, 05.12.2006, p. 2293-2302.

Research output: Contribution to journalArticle

Tao, L, Mogilner, A, Civelekoglu-Scholey, G, Wollman, R, Evans, J, Stahlberg, H & Scholey, JM 2006, 'A Homotetrameric Kinesin-5, KLP61F, Bundles Microtubules and Antagonizes Ncd in Motility Assays', Current Biology, vol. 16, no. 23, pp. 2293-2302. https://doi.org/10.1016/j.cub.2006.09.064
Tao, Li ; Mogilner, Alex ; Civelekoglu-Scholey, Gul ; Wollman, Roy ; Evans, James ; Stahlberg, Henning ; Scholey, Jonathan M. / A Homotetrameric Kinesin-5, KLP61F, Bundles Microtubules and Antagonizes Ncd in Motility Assays. In: Current Biology. 2006 ; Vol. 16, No. 23. pp. 2293-2302.
@article{5389962e40244b448609c6f305f8f607,
title = "A Homotetrameric Kinesin-5, KLP61F, Bundles Microtubules and Antagonizes Ncd in Motility Assays",
abstract = "Background: Mitosis depends upon the cooperative action of multiple microtubule (MT)-based motors. Among these, a kinesin-5, KLP61F, and the kinesin-14, Ncd, are proposed to generate antagonistic-sliding forces that control the spacing of the spindle poles. We tested whether purified KLP61F homotetramers and Ncd homodimers can generate a force balance capable of maintaining a constant spindle length in Drosophila embryos. Results: Using fluorescence microscopy and cryo-EM, we observed that purified full-length, motorless, and tailless KLP61F tetramers (containing a tetramerization domain) and Ncd dimers can all cross-link MTs into bundles in MgATP. In multiple-motor motility assays, KLP61F and Ncd drive plus-end and minus-end MT sliding at 0.04 and 0.1 μm/s, respectively, but the motility of either motor is decreased by increasing the mole fraction of the other. At the {"}balance point,{"} the mean velocity was zero and MTs paused briefly and then oscillated, taking ∼0.3 μm excursions at ∼0.02 μm/s toward the MT plus end and then the minus end. Conclusions: The results, combined with quantitative analysis, suggest that these motors could act as mutual brakes to modulate the rate of pole-pole separation and could maintain a prometaphase spindle displaying small fluctuations in its steady-state length.",
keywords = "CELLBIO",
author = "Li Tao and Alex Mogilner and Gul Civelekoglu-Scholey and Roy Wollman and James Evans and Henning Stahlberg and Scholey, {Jonathan M.}",
year = "2006",
month = "12",
day = "5",
doi = "10.1016/j.cub.2006.09.064",
language = "English (US)",
volume = "16",
pages = "2293--2302",
journal = "Current Biology",
issn = "0960-9822",
publisher = "Cell Press",
number = "23",

}

TY - JOUR

T1 - A Homotetrameric Kinesin-5, KLP61F, Bundles Microtubules and Antagonizes Ncd in Motility Assays

AU - Tao, Li

AU - Mogilner, Alex

AU - Civelekoglu-Scholey, Gul

AU - Wollman, Roy

AU - Evans, James

AU - Stahlberg, Henning

AU - Scholey, Jonathan M.

PY - 2006/12/5

Y1 - 2006/12/5

N2 - Background: Mitosis depends upon the cooperative action of multiple microtubule (MT)-based motors. Among these, a kinesin-5, KLP61F, and the kinesin-14, Ncd, are proposed to generate antagonistic-sliding forces that control the spacing of the spindle poles. We tested whether purified KLP61F homotetramers and Ncd homodimers can generate a force balance capable of maintaining a constant spindle length in Drosophila embryos. Results: Using fluorescence microscopy and cryo-EM, we observed that purified full-length, motorless, and tailless KLP61F tetramers (containing a tetramerization domain) and Ncd dimers can all cross-link MTs into bundles in MgATP. In multiple-motor motility assays, KLP61F and Ncd drive plus-end and minus-end MT sliding at 0.04 and 0.1 μm/s, respectively, but the motility of either motor is decreased by increasing the mole fraction of the other. At the "balance point," the mean velocity was zero and MTs paused briefly and then oscillated, taking ∼0.3 μm excursions at ∼0.02 μm/s toward the MT plus end and then the minus end. Conclusions: The results, combined with quantitative analysis, suggest that these motors could act as mutual brakes to modulate the rate of pole-pole separation and could maintain a prometaphase spindle displaying small fluctuations in its steady-state length.

AB - Background: Mitosis depends upon the cooperative action of multiple microtubule (MT)-based motors. Among these, a kinesin-5, KLP61F, and the kinesin-14, Ncd, are proposed to generate antagonistic-sliding forces that control the spacing of the spindle poles. We tested whether purified KLP61F homotetramers and Ncd homodimers can generate a force balance capable of maintaining a constant spindle length in Drosophila embryos. Results: Using fluorescence microscopy and cryo-EM, we observed that purified full-length, motorless, and tailless KLP61F tetramers (containing a tetramerization domain) and Ncd dimers can all cross-link MTs into bundles in MgATP. In multiple-motor motility assays, KLP61F and Ncd drive plus-end and minus-end MT sliding at 0.04 and 0.1 μm/s, respectively, but the motility of either motor is decreased by increasing the mole fraction of the other. At the "balance point," the mean velocity was zero and MTs paused briefly and then oscillated, taking ∼0.3 μm excursions at ∼0.02 μm/s toward the MT plus end and then the minus end. Conclusions: The results, combined with quantitative analysis, suggest that these motors could act as mutual brakes to modulate the rate of pole-pole separation and could maintain a prometaphase spindle displaying small fluctuations in its steady-state length.

KW - CELLBIO

UR - http://www.scopus.com/inward/record.url?scp=33751421228&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33751421228&partnerID=8YFLogxK

U2 - 10.1016/j.cub.2006.09.064

DO - 10.1016/j.cub.2006.09.064

M3 - Article

C2 - 17141610

AN - SCOPUS:33751421228

VL - 16

SP - 2293

EP - 2302

JO - Current Biology

JF - Current Biology

SN - 0960-9822

IS - 23

ER -