Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Kinesin’s tail domain is an inhibitory regulator of the motor domain

Nature Cell Biology volume 1, pages 288–292 (1999)Cite this article

Abstract

When not bound to cargo, the motor protein kinesin is in an inhibited state that has low microtubule-stimulated ATPase activity. Inhibition serves to minimize the dissipation of ATP and to prevent mislocalization of kinesin in the cell. Here we show that this inhibition is relieved when kinesin binds to an artificial cargo. Inhibition is mediated by kinesin’s tail domain: deletion of the tail activates the ATPase without need of cargo binding, and inhibition is re-established by addition of exogenous tail peptide. Both ATPase and motility assays indicate that the tail does not prevent kinesin from binding to microtubules, but rather reduces the motor’s stepping rate.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Structural model for kinesin and its regulation.
Figure 2: Coomassie-stained SDS–polyacrylamide gels of wild-type and mutant kinesin proteins.
Figure 3: Kinesin’s ATPase rate is activated by cargo binding or by deletion of its tail domain.
Figure 4: Exogenous kinesin tail protein inhibits kinesin’s ATPase activity and motility.
Figure 5: A structural model of tail-mediated inhibition.

Similar content being viewed by others

References

  1. Hurd, D. D. & Saxton, W. M. Kinesin mutations cause motor neuron disease phenotypes by disrupting fast axonal transport in Drosophila. Genetics 144, 1075–1085 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Coy, D. L. & Howard, J. Organelle transport and sorting in axons. Curr. Opin. Neurobiol. 4, 662–667 (1994).

    Article  CAS  Google Scholar 

  3. Bloom, G. S. & Endow, S. A. Motor proteins I: kinesins. Protein Profile 2, 1109–1171 (1995).

    CAS  Google Scholar 

  4. Hirokawa, N. Kinesin and dynein superfamily proteins and the mechanism of organelle transport. Science 279, 519–526 (1998).

    Article  CAS  Google Scholar 

  5. Howard, J. Molecular motors: structural adaptations to cellular functions. Nature 389, 561–567 (1997).

    Article  CAS  Google Scholar 

  6. Hollenbeck, P. J. The distribution, abundance and subcellular localization of kinesin. J. Cell Biol. 108, 2335–2342 (1989).

    Article  CAS  Google Scholar 

  7. Coy, D. L., Wagenbach, M. & Howard, J. Kinesin takes one eight-nanometer step for each ATP that it hydrolyzes. J. Biol. Chem. 276, 3667–3671 (1999).

    Article  Google Scholar 

  8. Berne, R. M. & Levy, M. N. Physiology 3rd edn (Mosby, St Louis, 1993).

    Google Scholar 

  9. Saxton, W. M. et al. Drosophila kinesin: characterization of microtubule motility and ATPase. Proc. Natl Acad. Sci. USA 85, 1109–1113 (1988).

    Article  CAS  Google Scholar 

  10. Hackney, D. D., Levitt, J. D. & Wagner, D. D. Characterization of alpha 2 beta 2 and alpha 2 forms of kinesin. Biochem. Biophys. Res. Commun. 174, 810–815 (1991).

    Article  CAS  Google Scholar 

  11. Yang, J. T., Laymon, R. A. & Goldstein, L. S. A three-domain structure of kinesin heavy chain revealed by DNA sequence and microtubule binding analyses. Cell 56, 879–889 (1989).

    Article  CAS  Google Scholar 

  12. de Cuevas, M., Tao, T. & Goldstein, L. S. Evidence that the stalk of Drosophila kinesin heavy chain is an alpha-helical coiled coil. J. Cell Biol. 116, 957–965 (1992).

    Article  CAS  Google Scholar 

  13. Huang, T. G., Suhan, J. & Hackney, D. D. Drosophila kinesin motor domain extending to amino acid position 392 is dimeric when expressed in Escherichia coli. J. Biol. Chem. 269, 16502–16507 (1994).

    CAS  PubMed  Google Scholar 

  14. Kozielski, F. et al. The crystal structure of dimeric kinesin and implications for microtubule-dependent motility. Cell 91, 985–994 (1997).

    Article  CAS  Google Scholar 

  15. Kull, F. J., Sablin, E. P., Lau, R., Fletterick, R. J. & Vale, R. D. Crystal structure of the kinesin motor domain reveals a structural similarity to myosin. Nature 380, 550–555 (1996).

    Article  CAS  Google Scholar 

  16. Yang, J. T., Saxton, W. M., Stewart, R. J., Raff, E. C. & Goldstein, L. S. Evidence that the head of kinesin is sufficient for force generation and motility in vitro. Science 249, 42–47 (1990).

    Article  CAS  Google Scholar 

  17. Cyr, J. L., Pfister, K. K., Bloom, G. S., Slaughter, C. A. & Brady, S. T. Molecular genetics of kinesin light chains: generation of isoforms by alternative splicing. Proc. Natl Acad. Sci. USA 88, 10114–10118 (1991).

    Article  CAS  Google Scholar 

  18. Gauger, A. K. & Goldstein, L. S. The Drosophila kinesin light chain. Primary structure and interaction with kinesin heavy chain. J. Biol. Chem. 268, 13657–13666 (1993).

    CAS  PubMed  Google Scholar 

  19. Verhey, K. J. et al. Light chain-dependent regulation of kinesin’s interaction with microtubules. J. Cell. Biol. 143, 1053–1066 (1998).

    Article  CAS  Google Scholar 

  20. Skoufias, D. A., Cole, D. G., Wedaman, K. P. & Scholey, J. M. The carboxyl-terminal domain of kinesin heavy chain is important for membrane binding. J. Biol. Chem. 269, 1477–1485 (1994).

    CAS  PubMed  Google Scholar 

  21. Bi, G. Q. et al. Kinesin- and myosin-driven steps of vesicle recruitment for Ca2+-regulated exocytosis. J. Cell. Biol. 138, 999–1008 (1997).

    Article  CAS  Google Scholar 

  22. Hackney, D. D., Levitt, J. D. & Suhan, J. Kinesin undergoes a 9 S to 6 S conformational transition. J. Biol. Chem. 267, 8696–8701 (1992).

    CAS  PubMed  Google Scholar 

  23. Trybus, K. M., Huiatt, T. W. & Lowey, S. A bent monomeric conformation of myosin from smooth muscle. Proc. Natl Acad. Sci. USA 79, 6151–6155 (1982).

    Article  CAS  Google Scholar 

  24. Trybus, K. M., Freyzon, Y., Faust, L. Z. & Sweeney, H. L. Spare the rod, spoil the regulation: necessity for a myosin rod. Proc. Natl Acad. Sci. USA 94, 48–52 (1997).

    Article  CAS  Google Scholar 

  25. Kuznetsov, S. A., Vaisberg, Y. A., Rothwell, S. W., Murphy, D. B. & Gelfand, V. I. Isolation of a 45-kDa fragment from the kinesin heavy chain with enhanced ATPase and microtubule-binding activities. J. Biol. Chem. 264, 589–595 (1989).

    CAS  PubMed  Google Scholar 

  26. Stewart, R. J., Thaler, J. P. & Goldstein, L. S. Direction of microtubule movement is an intrinsic property of the motor domains of kinesin heavy chain and Drosophila ncd protein. Proc. Natl Acad. Sci. USA 90, 5209–5213 (1993).

    Article  CAS  Google Scholar 

  27. Jiang, M. Y. & Sheetz, M. P. Cargo-activated ATPase activity of kinesin. Biophys. J. 68 (suppl.), 283–284 (1995).

    Google Scholar 

  28. Lockhart, A., Crevel, I. M. T. C. & Cross, R. A. Kinesin and ncd bind through a single head to microtubules and compete for a shared MT binding site. J. Mol. Biol. 249, 763–771 (1995).

    Article  CAS  Google Scholar 

  29. Amos, L. Kinesin from pig brain studied by electron microscopy. J. Cell Sci. 87, 105–111 (1987).

    CAS  PubMed  Google Scholar 

  30. Hirokawa, N. et al. Submolecular domains of bovine brain kinesin identified by electron microscopy and monoclonal antibody decoration. Cell 56, 867–878 (1989).

    Article  CAS  Google Scholar 

  31. Lupas, A. Coiled coils: new structures and new functions. Trends Biochem. Sci. 21, 375–382 (1996).

    Article  CAS  Google Scholar 

  32. Berger, B. et al. Predicting coiled coils by use of pairwise residue correlations. Proc. Natl Acad. Sci. USA 92, 8259–8263 (1995).

    Article  CAS  Google Scholar 

  33. Hancock, W. O. & Howard, J. Processivity of the motor protein kinesin requires two heads. J. Cell Biol. 140, 1395–1405 (1998).

    Article  CAS  Google Scholar 

  34. Bloomfield, V., Dalton, W. O. & Van Holde, K. E. Frictional coefficients of multisubunit structures. I. Theory. Biopolymers 5, 135–148 (1967).

    Article  CAS  Google Scholar 

  35. Fersht, A. Enzyme Structure and Mechanism (W.H. Freeman, New York, 1985).

    Google Scholar 

  36. Hollenbeck, P. J. Phosphorylation of neuronal kinesin heavy and light chains in vivo. J. Neurochem. 60, 2265–2275 (1993).

    Article  CAS  Google Scholar 

  37. Lee, K. D. & Hollenbeck, P. J. Phosphorylation of kinesin in vivo correlates with organelle association and neurite outgrowth. J. Biol. Chem. 264, 5600–5605 (1995).

    Article  Google Scholar 

  38. Howard, J., Hunt, A. J. & Baek, S. Assay of microtubule movement driven by single kinesin molecules. Methods Cell Biol. 39, 137–147 (1993).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank A. Hunter, P. Detwiler, E. Lumpkin and R. Sawhney for comments on an earlier version of this manuscript. This work was supported by NIH grant AR40593 (to J.H.). D.L.C. was supported by NIH Molecular Biophysics Training Grant GM08268 and by the Achievement Reward for College Scientists.

Correspondence and requests for materials should be addressed to J.H. The vectors encoding the following proteins have been deposited at GenBank under the indicated accession numbers: vector pPK113 (kinesin α2), AF053733; pPK121 (kinesin α2β2), AF055298; pPK115 (Δhinge), AF117643; pH911 (tail-911), AF117644; pPK124 (Δtail), AF161077; p864INS (tail-864), AF116269.

Author information

Authors and Affiliations

  1. Department of Physiology & Biophysics, University of Washington, Box 357290, Seattle, 98195-7290, Washington, USA

    David L. Coy, William O. Hancock, Michael Wagenbach & Jonathon Howard

Authors
  1. David L. Coy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. William O. Hancock
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michael Wagenbach
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jonathon Howard
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jonathon Howard.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Coy, D., Hancock, W., Wagenbach, M. et al. Kinesin’s tail domain is an inhibitory regulator of the motor domain. Nat Cell Biol 1, 288–292 (1999). https://doi.org/10.1038/13001

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/13001

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing