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.

  • Letter
  • Published:

A microtubule-binding domain in dynactin increases dynein processivity by skating along microtubules

Nature Cell Biology volume 8, pages 264–270 (2006)Cite this article

Abstract

Microtubule-associated proteins (MAPs) use particular microtubule-binding domains that allow them to interact with microtubules in a manner specific to their individual cellular functions. Here, we have identified a highly basic microtubule-binding domain in the p150 subunit of dynactin that is only present in the dynactin members of the CAP–Gly family of proteins. Using single-particle microtubule-binding assays, we found that the basic domain of dynactin moves progressively along microtubules in the absence of molecular motors — a process we term 'skating'. In contrast, the previously described CAP–Gly domain of dynactin remains firmly attached to a single point on microtubules. Further analyses showed that microtubule skating is a form of one-dimensional diffusion along the microtubule. To determine the cellular function of the skating phenomenon, dynein and the dynactin microtubule-binding domains were examined in single-molecule motility assays. We found that the basic domain increased dynein processivity fourfold whereas the CAP–Gly domain inhibited dynein motility. Our data show that the ability of the basic domain of dynactin to skate along microtubules is used by dynein to maintain longer interactions for each encounter with microtubules.

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: Dynactin contains two microtubule-binding domains.
Figure 2: Single-particle behaviour of protein-coated beads.
Figure 3: Quantitative analyses of microtubule interactions.

Similar content being viewed by others

References

  1. Karki, S. & Holzbaur, E. L. Affinity chromatography demonstrates a direct binding between cytoplasmic dynein and the dynactin complex. J. Biol. Chem. 270, 28806–28811 (1995).

    Article  CAS  Google Scholar 

  2. Vaughan, K. T. & Vallee, R. B. Cytoplasmic dynein binds dynactin through a direct interaction between the intermediate chains and p150Glued. J. Cell Biol. 131, 1507–1516 (1995).

    Article  CAS  Google Scholar 

  3. Vaughan, P. S., Leszyk, J. D. & Vaughan, K. T. Cytoplasmic dynein intermediate chain phosphorylation regulates binding to dynactin. J. Biol. Chem. 276, 26171–26179 (2001).

    Article  CAS  Google Scholar 

  4. King, S. J. et al. Analysis of the dynein-dynactin interaction in vitro and in vivo. Mol. Biol. Cell 14, 5089–5097 (2003).

    Article  CAS  Google Scholar 

  5. Karki, S. & Holzbaur, E. L. Cytoplasmic dynein and dynactin in cell division and intracellular transport. Curr. Opin. Cell Biol. 11, 45–53 (1999).

    Article  CAS  Google Scholar 

  6. Reiner, O. et al. Isolation of a Miller–Dieker lissencephaly gene containing G protein β-subunit-like repeats. Nature 364, 717–721 (1993).

    Article  CAS  Google Scholar 

  7. LaMonte, B. H. et al. Disruption of dynein–dynactin inhibits axonal transport in motor neurons causing late-onset progressive degeneration. Neuron 34, 715–727 (2002).

    Article  CAS  Google Scholar 

  8. Hafezparast, M. et al. Mutations in dynein link motor neuron degeneration to defects in retrograde transport. Science 300, 808–812 (2003).

    Article  CAS  Google Scholar 

  9. Puls, I. et al. Mutant dynactin in motor neuron disease. Nature Genet. 33, 455–456 (2003).

    Article  CAS  Google Scholar 

  10. Scheel, J. et al. Purification and analysis of authentic CLIP-170 and recombinant fragments. J. Biol. Chem. 274, 25883–25891 (1999).

    Article  CAS  Google Scholar 

  11. Hirokawa, N., Shiomura, Y. & Okabe, S. Tau proteins: the molecular structure and mode of binding on microtubules. J. Cell Biol. 107, 1449–1459 (1988).

    Article  CAS  Google Scholar 

  12. Wang, Z. & Sheetz, M. P. One-dimensional diffusion on microtubules of particles coated with cytoplasmic dynein and immunoglobulins. Cell Struct. Funct. 24, 373–383 (1999).

    Article  CAS  Google Scholar 

  13. Vale, R. D., Soll, D. R. & Gibbons, I. R. One-dimensional diffusion of microtubules bound to flagellar dynein. Cell 59, 915–925 (1989).

    Article  CAS  Google Scholar 

  14. King, S. J. & Schroer, T. A. Dynactin increases the processivity of the cytoplasmic dynein motor. Nature Cell Biol. 2, 20–24 (2000).

    Article  CAS  Google Scholar 

  15. Quintyne, N. J. et al. Dynactin is required for microtubule anchoring at centrosomes. J. Cell Biol. 147, 321–334 (1999).

    Article  CAS  Google Scholar 

  16. Sloboda, R. D. & Rosenbaum, J. L. Purification and assay of microtubule-associated proteins (MAPs). Methods Enzymol. 85, 409–416 (1982).

    Article  CAS  Google Scholar 

  17. Bingham, J. B., King, S. J. & Schroer, T. A. Purification of dynactin and dynein from brain tissue. Methods Enzymol. 298, 171–184 (1998).

    Article  CAS  Google Scholar 

  18. Mallik, R. et al. Building complexity: an in vitro study of cytoplasmic dynein with in vivo implications. Curr. Biol. 15, 2075–2085 (2005).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank G. J. Wyckoff for initial sequence alignments, R. Mallik and S. Gross for the MSD program and helpful discussions, H. Goodson for the CLIP-170 H1 polypeptides, H. Paudel for the full-length tau construct, and L. Ehler for critical reading of the manuscript. This work was supported by a National Institutes of Health (NIH) award NS48501 (S.J.K.) and in part by a University of Missouri-Kansas City SEARCH Undergraduate Research award (T.L.C.).

Author information

Author notes

  1. Nicholas J. Quintyne

    Present address: Wilkes Honors College of Florida Atlantic University, WB 217, 5353 Parkside Drive, Jupiter, FL 33458, USA

Authors and Affiliations

  1. Division of Molecular Biology and Biochemistry, School of Biological Sciences, University of Missouri-Kansas City, 5007 Rockhill Rd., Kansas City, 64110, MO, USA

    Tara L. Culver–Hanlon, Stephanie A. Lex, Andrew D. Stephens & Stephen J. King

  2. Department of Biological Sciences, University of Pittsburgh, 4249 Fifth Avenue, Pittsburgh, 15260, PA, USA

    Nicholas J. Quintyne

Authors
  1. Tara L. Culver–Hanlon
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Stephanie A. Lex
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Andrew D. Stephens
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nicholas J. Quintyne
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Stephen J. King
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Stephen J. King.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary figure S1 and S2 plus Supplementary Information and Supplementary Methods (PDF 408 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Culver–Hanlon, T., Lex, S., Stephens, A. et al. A microtubule-binding domain in dynactin increases dynein processivity by skating along microtubules. Nat Cell Biol 8, 264–270 (2006). https://doi.org/10.1038/ncb1370

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb1370

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