Skip to main content
SpringerLink
Log in

A torque component present in mitotic kinesin Eg5 revealed by three-dimensional tracking

  • Brief Communication
  • Published:

From Nature Structural & Molecular Biology

View current issue Submit your manuscript

Abstract

Mitotic kinesin Eg5 is a homotetrameric molecular motor that cross-links and slides microtubules. The extent to which Eg5 moves processively is not clear. Here we use three-dimensional tracking of a quantum dot attached to the microtubule in a motility assay to directly visualize the corkscrew motion of a sliding microtubule. We show that the rotational pitch of microtubule sliding conveniently reports on the processivity of the driving motors, confirming that two-headed Eg5 is much less processive than two-headed kinesin-1.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1: Observation of the corkscrew motion of a sliding microtubule driven by Eg5 under a three-dimensional tracking microscope.
Figure 2: Determination of the pitch and direction of rotation.

Similar content being viewed by others

References

  1. Sawin, K.E., LeGuellec, K., Philippe, M. & Mitchison, T.J. Nature 359, 540–543 (1992).

    Article  CAS  Google Scholar 

  2. Blangy, A. et al. Cell 83, 1159–1169 (1995).

    Article  CAS  Google Scholar 

  3. Valentine, M.T., Fordyce, P.M., Krzysiak, T.C., Gilbert, S.P. & Block, S.M. Nat. Cell Biol. 8, 470–476 (2006).

    Article  CAS  Google Scholar 

  4. Kaseda, K., Crevel, I., Hirose, K. & Cross, R.A. EMBO Rep. 9, 761–765 (2008).

    Article  CAS  Google Scholar 

  5. Howard, J., Hudspeth, A.J. & Vale, R.D. Nature 342, 154–158 (1989).

    Article  CAS  Google Scholar 

  6. Vale, R.D. et al. Nature 380, 451–453 (1996).

    Article  CAS  Google Scholar 

  7. Block, S.M., Goldstein, L.S. & Schnapp, B.J. Nature 348, 348–352 (1990).

    Article  CAS  Google Scholar 

  8. Ray, S., Meyhöfer, E., Milligan, R.A. & Howard, J. J. Cell Biol. 121, 1083–1093 (1993).

    Article  CAS  Google Scholar 

  9. Yajima, J. & Cross, R.A. Nat. Chem. Biol. 1, 338–341 (2005).

    Article  CAS  Google Scholar 

  10. Walker, R.A., Salmon, E.D. & Endow, S.A. Nature 347, 780–782 (1990).

    Article  CAS  Google Scholar 

  11. Okada, Y., Higuchi, H. & Hirokawa, N. Nature 424, 574–577 (2003).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank D.R. Drummond and R.A. Cross for critical comments and discussion on the manuscript, T. Usui (University of Tsukuba) for the Eg5 clone, T. Masaike and I. Terashima for supporting the experiments, and J. Ohkawa and K. Matsui for the microscope technique. This work was supported in part by Grants-in-Aid for Young Scientists (No. 20710170) to J.Y. and for Scientific Research on Priority Areas (No. 509 and 521) to T.N. from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and by the grant to T.N. from the New Energy and Industrial Technology Development Organization (NEDO).

Author information

Author notes

  1. Junichiro Yajima and Kana Mizutani: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Physics, Gakushuin University, 1-5-1 Mejiro, Toshima-ku, Tokyo, 171-8588, Japan

    Junichiro Yajima, Kana Mizutani & Takayuki Nishizaka

Authors
  1. Junichiro Yajima
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kana Mizutani
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Takayuki Nishizaka
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

J.Y. and K.M. contributed equally to this work: K.M. designed and developed the new microscopy method; J.Y. conceived the experiments, performed rotation measurements and the data analyses and wrote the manuscript. T.N. supervised this work.

Corresponding author

Correspondence to Takayuki Nishizaka.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4, Supplementary Methods (PDF 13065 kb)

Supplementary Video 1

First ∼1 second shows a sliding rhodamine-labeled microtubule (0.7 s intervals, ×10 actual speed, rhodamine filter set, Nikon). Remaining ∼9 seconds shows a moving QD 525 (Invitrogen) fixed to the microtubule via a streptavidin-biotin interaction (2.8 s intervals ×300 actual speed, Qdot 525 filter set, Chroma). Image, 14 μm high × 52 μm wide. (MOV 751 kb)

Supplementary Video 2

The viewing angle is changed every 10 degrees by 360 degrees along a longitudinal axis of the microtubule. The colors (blue, sky blue, green, yellow) of the dots show the height of z-position. The blue is lowest and the yellow is highest. A distance between blue dot and yellow dot is ∼70 nm. See Fig. 1c (MOV 1594 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yajima, J., Mizutani, K. & Nishizaka, T. A torque component present in mitotic kinesin Eg5 revealed by three-dimensional tracking. Nat Struct Mol Biol 15, 1119–1121 (2008). https://doi.org/10.1038/nsmb.1491

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nsmb.1491

  • Springer Nature America, Inc.

This article is cited by

Search

Navigation