Abstract
A polarity complex of PAR-3, PAR-6 and atypical protein kinase C (aPKC) functions in various cell-polarization events, including neuron specification1,2,3,4. The small GTPase Cdc42 binds to PAR-6 and regulates cell polarity. However, little is known about the downstream signals of the Cdc42–PAR protein complex. Here, we found that PAR-3 directly interacted with STEF/Tiam1, which are Rac-specific guanine nucleotide-exchange factors, and that STEF formed a complex with PAR-3–aPKC–PAR-6–Cdc42-GTP. Cdc42 induces lamellipodia in a Rac-dependent manner in N1E-115 neuroblastoma cells. Disruption of Cdc42–PAR-6 or PAR-3–STEF binding inhibited Cdc42-induced lamellipodia but not filopodia. The isolated STEF-binding PAR-3 fragment was sufficient to induce lamellipodia independently of Cdc42 and PAR-6. PAR-3 is required for Cdc42-induced Rac activation, but is not essential for lamellipodia formation itself. In cultured hippocampal neurons, STEF accumulated at the tip of the growing axon and colocalized with PAR-3. The spatio-temporal activation and signalling of Cdc42–PAR-6–PAR-3–STEF/Tiam1–Rac seem to be involved in neurite growth and axon specification. We propose that the PAR-6–PAR-3 complex mediates Cdc42-induced Rac activation by means of STEF/Tiam1, and that this process seems to be required for the establishment of neuronal polarity.
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
Ohno, S. Intercellular junctions and cellular polarity: the PAR–aPKC complex, a conserved core cassette playing fundamental roles in cell polarity. Curr. Opin. Cell Biol. 13, 641–648 (2001).
Macara, I. G. Parsing the polarity code. Nature Rev. Mol. Cell Biol. 5, 220–231 (2004).
Shi, S. H., Jan, L. Y. & Jan, Y. N. Hippocampal neuronal polarity specified by spatially localized mPar3/mPar6 and PI 3-kinase activity. Cell 112, 63–75 (2003).
Nishimura, T. et al. Role of the PAR-3–KIF3 complex in the establishment of neuronal polarity. Nature Cell Biol. 6, 328–334 (2004).
Craig, A. M. & Banker, G. Neuronal polarity. Annu. Rev. Neurosci. 17, 267–310 (1994).
Bradke, F. & Dotti, C. G. Establishment of neuronal polarity: lessons from cultured hippocampal neurons. Curr. Opin. Neurobiol. 10, 574–581 (2000).
Fukata, Y., Kimura, T. & Kaibuchi, K. Axon specification in hippocampal neurons. Neurosci. Res. 43, 305–315 (2002).
Kaibuchi, K., Kuroda, S. & Amano, M. Regulation of the cytoskeleton and cell adhesion by the Rho family GTPases in mammalian cells. Annu. Rev. Biochem. 68, 459–486 (1999).
Burridge, K. & Wennerberg, K. Rho and Rac take center stage. Cell 116, 167–179 (2004).
Etienne-Manneville, S. & Hall, A. Rho GTPases in cell biology. Nature 420, 629–635 (2002).
Schwamborn, J. C. & Puschel, A. W. The sequential activity of the GTPases Rap1B and Cdc42 determines neuronal polarity. Nature Neurosci. 7, 923–929 (2004).
Menager, C., Arimura, N., Fukata, Y. & Kaibuchi, K. PIP3 is involved in neuronal polarization and axon formation. J. Neurochem. 89, 109–118 (2004).
Etienne-Manneville, S. & Hall, A. Integrin-mediated activation of Cdc42 controls cell polarity in migrating astrocytes through PKCζ. Cell 106, 489–498 (2001).
Yamanaka, T. et al. PAR-6 regulates aPKC activity in a novel way and mediates cell–cell contact-induced formation of the epithelial junctional complex. Genes Cells 6, 721–731 (2001).
Leeuwen, F. N. et al. The guanine nucleotide exchange factor Tiam1 affects neuronal morphology: opposing roles for the small GTPases Rac and Rho. J. Cell Biol. 139, 797–807 (1997).
Matsuo, N., Terao, M., Nabeshima, Y. & Hoshino, M. Roles of STEF/Tiam1, guanine nucleotide exchange factors for Rac1, in regulation of growth cone morphology. Mol. Cell. Neurosci. 24, 69–81 (2003).
Kunda, P., Paglini, G., Quiroga, S., Kosik, K. & Caceres, A. Evidence for the involvement of Tiam1 in axon formation. J. Neurosci. 21, 2361–2372 (2001).
Kozma, R., Sarner, S., Ahmed, S. & Lim, L. Rho family GTPases and neuronal growth cone remodeling: relationship between increased complexity induced by Cdc42Hs, Rac1, and acetylcholine and collapse induced by RhoA and lysophosphatidic acid. Mol. Cell. Biol. 17, 1201–1211 (1997).
Matsuo, N., Hoshino, M., Yoshizawa, M. & Nabeshima, Y. Characterization of STEF, a guanine nucleotide exchange factor for Rac1, required for neurite growth. J. Biol.Chem. 277, 2860–2868 (2002).
Lin, D. et al. A mammalian PAR-3–PAR-6 complex implicated in Cdc42/Rac1 and aPKC signaling and cell polarity. Nature Cell Biol. 2, 540–547 (2000).
Michiels, F. et al. Regulated membrane localization of Tiam1, mediated by the NH2-terminal pleckstrin homology domain, is required for Rac-dependent membrane ruffling and C-Jun NH2-terminal kinase activation. J. Cell Biol. 137, 387–398 (1997).
Mertens, A. E., Roovers, R. C. & Collard, J. G. Regulation of Tiam1–Rac signaling. FEBS Lett. 546, 11–16 (2003).
Fleming, I. N., Gray, A. & Downes, C. P. Regulation of the Rac1-specific exchange factor Tiam1 involves both phosphoinositide 3-kinase-dependent and -independent components. Biochem. J. 351, 173–182 (2000).
Rolls, M. M. & Doe, C. Q. Baz, Par-6 and aPKC are not required for axon or dendrite specification in Drosophila. Nature Neurosci. 7, 1293–1295 (2004).
Mishima, A. et al. Over-expression of PAR-3 suppresses contact-mediated inhibition of cell migration in MDCK cells. Genes Cells 7, 581–596 (2002).
Habets, G. G. et al. Identification of an invasion-inducing gene, Tiam1, that encodes a protein with homology to GDP–GTP exchangers for Rho-like proteins. Cell 77, 537–549 (1994).
Hordijk, P. L. et al. Inhibition of invasion of epithelial cells by Tiam1–Rac signaling. Science 278, 1464–1466 (1997).
Sander, E. E. et al. Matrix-dependent Tiam1/Rac signaling in epithelial cells promotes either cell–cell adhesion or cell migration and is regulated by phosphatidylinositol 3-kinase. J. Cell Biol. 143, 1385–1398 (1998).
Hoshino, M. et al. Identification of the stef gene that encodes a novel guanine nucleotide exchange factor specific for Rac1. J. Biol. Chem. 274, 17837–17844 (1999).
Nishimura, T. et al. CRMP-2 regulates polarized Numb-mediated endocytosis for axon growth. Nature Cell Biol. 5, 819–826 (2003).
Acknowledgements
We thank members of the Kaibuchi lab for useful discussions and for preparing some materials; N. Ui and M. Yoshizaki for technical assistance; and T. Ishii for secretarial assistance. This research was supported, in part, by Grant-in-Aid for Scientific Research; Grant-in-Aid for Creative Scientific Research; The 21st Century Center of Excellence Program from the Ministry of Education, Culture, Sports, Science and Technology of Japan; The Research Grant for Nervous and Mental Disorders from the Ministry of Health, Labour and Welfare; and The Pharmaceuticals and Medical Devices Agency. T.N. is a Research Fellow of the Japan Society for the Promotion of Science.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary information
Supplementary figures S1, S2, S3, S4 and S5 (PDF 959 kb)
Rights and permissions
About this article
Cite this article
Nishimura, T., Yamaguchi, T., Kato, K. et al. PAR-6–PAR-3 mediates Cdc42-induced Rac activation through the Rac GEFs STEF/Tiam1. Nat Cell Biol 7, 270–277 (2005). https://doi.org/10.1038/ncb1227
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ncb1227
This article is cited by
-
Advances in Understanding the Molecular Mechanisms of Neuronal Polarity
Molecular Neurobiology (2023)
-
Apical–basal polarity and the control of epithelial form and function
Nature Reviews Molecular Cell Biology (2022)
-
Rho–Rho-Kinase Regulates Ras-ERK Signaling Through SynGAP1 for Dendritic Spine Morphology
Neurochemical Research (2022)
-
The polarity protein PARD3 and cancer
Oncogene (2021)
-
Manipulating oligodendrocyte intrinsic regeneration mechanism to promote remyelination
Cellular and Molecular Life Sciences (2021)