Abstract
Cytoplasmic dynein supports long-range intracellular movements of cargo in vivo but does not appear to be a processive motor protein by itself. We show here that the dynein activator, dynactin, binds microtubules and increases the average length of cytoplasmic-dynein-driven movements without affecting the velocity or microtubule-stimulated ATPase kinetics of cytoplasmic dynein. Enhancement of microtubule binding and motility by dynactin are both inhibited by an antibody to dynactin’s microtubule-binding domain. These results indicate that dynactin acts as a processivity factor for cytoplasmic-dynein-based motility and provide the first evidence that cytoskeletal motor processivity can be affected by extrinsic factors.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Finer, J. T., Simmons, R. M. & Spudich, J. A. Single myosin molecule mechanics: piconewton forces and nanometre steps. Nature 368, 113–119 (1994).
Shingyoji, C., Higuchi, H., Yoshimura, M., Katayama, E. & Yanagida, T. Dynein arms are oscillating force generators. Nature 393, 711–714 (1998).
Howard, J., Hudspeth, A. J. & Vale, R. D. Movement of microtubules by single kinesin molecules. Nature 342, 154–158 (1989).
Block, S. M., Goldstein, L. B. & Schnapp, B. J. Bead movement by single kinesin molecules studied with optical tweezers. Nature 348, 348–352 (1990).
Hackney, D. D. The kinetic cycles of myosin, kinesin, and dynein. Annu. Rev. Physiol. 58, 731–750 (1996).
Allan, V. Motor proteins: a dynamic duo. Curr. Biol. 6, 630–633 (1996).
Karki, S. & Holzbaur, E. L. F. Cytoplasmic dynein and dynactin in cell division and intracellular transport. Curr. Opin. Cell Biol. 11, 45–53 (1999).
Schroer, T. A. Structure and function of dynactin. Semin. Cell Dev. Biol. 7, 321–328 (1996).
Schafer, D. A., Gill, S. R., Cooper, J. A., Heuser, J. E. & Schroer, T. A. Ultrastructural analysis of the dynactin complex: an actin-related protein is a component of a filament that resembles f-actin. J. Cell Biol. 126, 403–412 (1994).
Holleran, E. A., Tokito, M. K., Karki, S. & Holzbaur, E. L. F. Centractin (Arp1) associates with spectrin revealing a potential mechanism to link dynactin to intracellular organelles. J. Cell Biol. 135, 1815–1829 (1996).
Eckley, D. M. et al. Analysis of dynactin subcomplexes reveals a novel actin-related protein associated with the Arp1 minifilament pointed end. J. Cell Biol. 147, 307–319 (1999).
Karki, S. & Holzbaur, E. L. F. Affinity chromatography demonstrates a direct binding between cytoplasmic dynein and the dynactin complex. J. Biol. Chem. 270, 28806–28811 (1995).
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).
Waterman-Storer, C. M., Karki, S. & Holzbaur, E. L. The p150Glued component of the dynactin complex binds to both microtubules and the actin-related protein centractin (Arp-1). Proc. Natl Acad. Sci. USA 92, 1634–1638 (1995).
Rickard, J. E. & Kreis, T. E. CLIPs for organelle-microtubule interactions. Trends Cell Biol. 6, 178–183 (1996).
Gill, S. R. et al. Dynactin, a conserved, ubiquitously expressed component of an activator of vesicle motility mediated by cytoplasmic dynein. J. Cell Biol. 115, 1639–1650 (1991).
Quintyne, N. J. et al. Dynactin is required for microtubule anchoring at fibroblast centrosomes. J. Cell Biol. 147, 321–334 (1999).
Vallee, R. B. & Sheetz, M. P. Targeting of motor proteins. Science 271, 1539–1544 (1996).
Shpetner, H. S., Paschal, B. M. & Vallee, R. B. Characterization of the microtubule-activated ATPase of brain cytoplasmic dynein (MAP 1C). J. Cell Biol. 107, 1001–1009 (1988).
Shimizu, T., Toyoshima, Y. Y., Edamatsu, M. & Vale, R. D. Comparison of the motile and enzymatic properties of two microtubule minus-end-directed motors, ncd and cytoplasmic dynein. Biochemistry 34, 1575–1582 (1995).
Holzbaur, E. L. F. & Johnson, K. A. Microtubules accelerate ADP release by dynein. Biochemistry 28, 7010–7016 (1989).
Wang, Z., Khan, S. & Sheetz, M. P. Single cytoplasmic dynein molecule movements: characterization and comparison with kinesin. Biophys. J. 69, 2011–2023 (1995).
Okada, Y. & Hirokawa, N. A processive single-headed motor: kinesin superfamily protein KIF1A. Science 283, 1152–1157 (1999).
Takada, S. & Kamiya, R. Functional reconstitution of Chlamydomonas outer dynein arms from α-β and γ subunits: requirement of a third factor. J Cell Biol. 126, 737–745 (1994).
Huang, C. F., Chang, C. B., Huang, C. & Farrell, J. E. Jr M phase phosphorylation of cytoplasmic dynein intermediate chain and p150Glued. J. Biol. Chem. 274, 14262–14269 (1999).
Farshori, P. & Holzbaur, E. L. F. Dynactin phosphorylation is modulated in response to cellular effectors. Biochem. Biophys. Res. Commun. 232, 810–816 (1997).
Niclas, J., Allan, V. J. & Vale, R. D. Cell cycle regulation of dynein association with membranes modulates microtubule-based organelle transport. J. Cell Biol. 133, 585–593 (1996).
Schroer, T. A. & Sheetz, M. P. Two activators of microtubule-based vesicle transport. J. Cell Biol. 115, 1309–1318 (1991).
Bingham, J. B., King, S. J. & Schroer, T. A. Purification of dynactin and dynein from brain tissue. Methods Enzymol. 298, 171–184 (1998).
Sloboda, R. D. & Rosenbaum, J. L. Purification and assay of microtubule-associated proteins (MAPs). Methods Enzymol. 85, 409–416 (1982).
Huang, T.-G. & Hackney, D. D. Drosophila kinesin minimal motor domain expressed in Escherichia coli: purification and kinetic characterization. J. Biol. Chem. 269, 16493–16501 (1994).
Schnapp, B. J. Viewing single microtubules by video light microscopy. Methods Enzymol. 134, 561–573 (1986).
Sakakibara, H., Kojima, H., Sakai, Y., Katayama, E. & Oiwa, K. Inner-arm dynein c of Chlamydomonas flagella is a single-headed processive motor. Nature 400, 586–590 (1999).
Vallee, R. B., Wall, J. S., Paschal, B. M. & Shpetner, H. S. Microtubule-associated protein 1C from brain is a two-headed cytosolic dynein. Nature 332, 561–563 (1988).
Amos, L. A. Brain dynein crossbridges microtubules into bundles. J. Cell Sci. 93, 19–28 (1989).
Acknowledgements
We thank members of the Schroer laboratory for valuable discussions; and L. Ehler and the Schroer laboratory for critical reading of the manuscript. This work was supported by grants from the NIH to T.A.S (GM 44589) and to S.J.K. (F32 GM 19061).
Correspondence and requests for materials should be addressed to T.A.S.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
King, S., Schroer, T. Dynactin increases the processivity of the cytoplasmic dynein motor. Nat Cell Biol 2, 20–24 (2000). https://doi.org/10.1038/71338
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/71338
This article is cited by
-
Schizophrenia-associated dysbindin modulates axonal mitochondrial movement in cooperation with p150glued
Molecular Brain (2021)
-
Dynein harnesses active fluctuations of microtubules for faster movement
Nature Physics (2020)
-
Molecular evidence for adaptive evolution of olfactory-related genes in cervids
Genes & Genomics (2020)
-
Force sharing and force generation by two teams of elastically coupled molecular motors
Scientific Reports (2019)
-
Cargo adaptors regulate stepping and force generation of mammalian dynein–dynactin
Nature Chemical Biology (2019)