Abstract
Coordination of microtubules and the actin cytoskeleton is important in several types of cell movement. mDia1 is a member of the formin-homology family of proteins and an effector of the small GTPase Rho. It contains the Rho-binding domain in its amino terminus and two distinct regions of formin homology, FH1 in the middle and FH2 in the carboxy terminus. Here we show that expression of mDia1(ΔN3), an active mDia1 mutant containing the FH1 and FH2 regions without the Rho-binding domain, induces bipolar elongation of HeLa cells and aligns microtubules in parallel to F-actin bundles along the long axis of the cell. The cell elongation and microtubule alignment caused by this mutant is abolished by co-expression of an FH2-region fragment, and expression of mDia1(ΔN3) containing point mutations in the FH2 region causes an increase in the amount of disorganized F-actin without cell elongation and microtubule alignment. These results indicate that mDia1 may coordinate microtubules and F-actin through its FH2 and FH1 regions, respectively.
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
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Lauffenburger, D. A. & Horwitz, A. F. Cell migration: a physically integrated molecular process. Cell 84, 359–369 (1996).
Mitchison, T. J. & Cramer, L. P. Actin-based cell motility and cell locomotion. Cell 84, 371–379 (1996).
Vasiliev, J. M. Polarization of pseudopodial activities: cytoskeletal mechanisms. J. Cell Sci. 98, 1–4 ( 1991).
Nobes, C. D. & Hall, A. Rho, Rac, and Cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia. Cell 81 , 53–62 (1995).
Kozma, R., Ahmed, S., Best, A. & Lim, L. The Ras-related protein Cdc42Hs and bradykinin promote formation of peripheral actin microspikes and filopodia in Swiss 3T3 fibroblasts. Mol. Cell. Biol. 15, 1942–1952 (1995).
Ridley, A. J. et al. The small GTP-binding protein rac regulates growth factor-induced membrane ruffling. Cell 70, 401– 410 (1992).
Machesky, L. M. & Insall, R. H. Scar1 and the related Wiskott–Aldrich syndrome protein, WASP, regulate the actin cytoskeleton through the Arp2/3 complex. Curr. Biol. 8, 1347–1356 (1998).
Rohatgi, R. et al. The interaction between N-WASP and the Arp2/3 complex links Cdc42-dependent signals to actin assembly. Cell 97, 221–231 (1999).
Ridley, A. J. & Hall, A. The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors. Cell 70, 389–399 (1992).
Leung, T., Manser, E., Tan, L. & Lim, L. A novel serine/threonine kinase binding the Ras-related RhoA GTPase which translocates the kinase to peripheral membranes. J. Biol. Chem. 270, 29051–29054 (1995).
Ishizaki, T. et al. The small GTP-binding protein Rho binds to and activates a 160 kDa Ser/Thr protein kinase homologous to myotonic dystrophy kinase. EMBO J. 15, 1885–1893 ( 1996).
Matsui, T. et al. Rho-associated kinase, a novel serine/threonine kinase, as a putative target for small GTP binding protein Rho. EMBO J. 15, 2208–2216 (1996).
Kimura, K. et al. Regulation of myosin phosphatase by Rho and Rho-associated kinase (Rho-kinase). Science 273, 245– 248 (1996).
Burridge, K., Chrzanowaska-Wodnicka, M. & Zhong, C. Focal adhesion assembly. Trends Cell Biol. 7, 342–347 ( 1997).
Watanabe, N. et al. p140mDia, a mammalian homolog of Drosophila diaphanous, is a target protein for Rho small GTPase and is a ligand for profilin. EMBO J. 16, 3044–3056 ( 1997).
Watanabe, N. et al. Cooperation between mDia1 and ROCK in Rho-induced actin reorganization . Nature Cell Biol. 1, 136– 143 (1999).
Nobes C. D. & Hall, A. Rho GTPases control polarity, protrusion, and adhesion during cell movement. J. Cell Biol. 144, 1235–1244 (1999).
Allen, W. E., Zicha, D., Ridley, A. J. & Jones, G. E. A role for Cdc42 in macrophage chemotaxis. J. Cell Biol. 141, 1147–1157 (1998).
Uehata, M. et al. Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension. Nature 389, 990–994 (1997).
Niggli, V. Rho-kinase in human neutrophils: a role in signalling for myosin light chain phosphorylation and cell migration. FEBS Lett. 445, 69–72 (1999).
Adams, A. E. et al. CDC42 and CDC43, two additional genes involved in budding and the establishment of cell polarity in the yeast Saccharomyces cerevisiae . J. Cell Biol. 111, 131– 142 (1990).
Drgonova, J., Drgon, T., Roh, D-H. & Cabib, E. The GTP-binding protein Rho1p is required for cell cycle progression and polarization of the yeast cell. J. Cell Biol. 146, 373– 387 (1999).
Bershadsky, A. D. & Futerman, A. H. Disruption of the Golgi apparatus by brefeldin A blocks cell polarization and inhibits directed cell migration. Proc. Natl Acad. Sci. USA 91, 5686–5689 (1994).
Evangelista, M. et al. Bni1p, a yeast formin linking Cdc42p and the actin cytoskeleton during polarized morphogenesis. Science 276, 118–122 (1997).
Tanaka, K. & Takai, Y. Control of reorganization of the actin cytoskeleton by Rho family small GTP-binding proteins in yeast . Curr. Opin. Cell Biol. 10, 112– 116 (1998).
Lee, L. et al. Control of mitotic spindle position by the Saccharomyces cerevisiae formin Bni1p. J. Cell Biol. 144, 947 –961 (1999).
Miller, R. K., Matheos, D. & Rose, M. D. The cortical localization of the microtubule orientation protein, Kar9p, is dependent upon actin and proteins required for polarization. J. Cell Biol. 144, 963 –975 (1999).
Enomoto, T. Microtubule disruption induces the formation of actin stress fibers and focal adhesions in cultured cells: possible involvement of the Rho signal cascade . Cell Struct. Funct. 21, 317– 326 (1996).
Zhang, Q., Magnusson, M. K. & Mosher, D. F. Lysophosphatidic acid and microtubule-destabilizing agents stimulate fibronectin matrix assembly through Rho-dependent actin stress fiber formation and cell contraction. Mol. Biol. Cell 8, 1415–1425 (1997).
Kaverina, I., Rottner, K. & Small, J. V. Targeting, capture, and stabilization of microtubules at early focal adhesions. J. Cell Biol. 142, 181–190 (1998).
Kaverina, I., Krylyshkina, O. & Small, J. V. Microtubule targeting of substrate contacts promotes their relaxation and dissociation. J. Cell Biol. 146 , 1033–1043 (1999).
Sawin, K. E. & Nurse, P. Regulation of cell polarity by microtubules in fission yeast. J. Cell Biol. 142 , 457–471 (1998).
Mata, J. & Nurse, P. Tea1 and the microtubular cytoskeleton are important for generating global spatial order within the fission yeast cell. Cell 89, 939– 949 (1997).
Maekawa, M. et al. Signalling from Rho to the actin cytoskeleton through protein kinases ROCK and LIM-kinase. Science 285, 895–898 (1999).
Wasserman, S. FH proteins as cytoskeletal organizers. Trends Cell Biol. 8, 111–115 (1998).
Norman, J. C. et al. ARF1 mediates paxillin recruitment to focal adhesions and potentiates Rho-stimulated stress fiber formation in intact and permeabilized Swiss 3T3 fibroblasts. J. Cell Biol. 143, 1981–1995 (1998).
Smilenov, L. B. et al. Focal adhesion motility revealed in stationary fibroblasts . Science 286, 1172–1174 (1999).
Elbaum, M. et al. in Cell Behaviour: Control and Mechanism of Motility (eds Lackie, J. M., Dunn, G. A. & Jones, G. E.), 147– 172 (Portland, London, 1999).
Ishizaki, T. et al. p160ROCK, a Rho-associated coiled-coil forming protein kinase, works downstream of Rho and induces focal adhesions. FEBS Lett. 404, 118–124 ( 1997).
Acknowledgements
We thank N. Watanabe for the pEGFP–mDia1ΔN3 construct and for his initial observation of microtubule alignment in mDia-expressing cells; H. Bito for advice on live-cell imaging; and A. Fujita, M. Maekawa and K. Kimura for discussions. This work was supported by a Grant in Aid for Specially Promoted Research from the Ministry of Education, Science, Sports and Culture of Japan, a grant from the Human Frontier Science Program (to S.N.), and grants from the Searle Foundation and the Japan Foundation for Applied Enzymology (to T.I.).
Author information
Authors and Affiliations
Corresponding author
Supplementary information
Figure S1
Effect of BFA on the actin cytoskeleton and microtubules in cells expressing mDia1(ΔN3). (PDF 134 kb)
Rights and permissions
About this article
Cite this article
Ishizaki, T., Morishima, Y., Okamoto, M. et al. Coordination of microtubules and the actin cytoskeleton by the Rho effector mDia1. Nat Cell Biol 3, 8–14 (2001). https://doi.org/10.1038/35050598
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/35050598
This article is cited by
-
The formin INF2 in disease: progress from 10 years of research
Cellular and Molecular Life Sciences (2020)
-
Cytoplasmic sequestration of the RhoA effector mDiaphanous1 by Prohibitin2 promotes muscle differentiation
Scientific Reports (2019)
-
Caveolin-1α regulates primary cilium length by controlling RhoA GTPase activity
Scientific Reports (2019)
-
Heparan sulfate proteoglycan (HSPG) can take part in cell division: inside and outside
Cellular and Molecular Life Sciences (2019)
-
Coordination of microtubule acetylation and the actin cytoskeleton by formins
Cellular and Molecular Life Sciences (2018)