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:

Direct interaction of two polarity complexes implicated in epithelial tight junction assembly

Nature Cell Biology volume 5, pages 137–142 (2003)Cite this article

Abstract

Tight junctions help establish polarity in mammalian epithelia by forming a physical barrier that separates apical and basolateral membranes. Two evolutionarily conserved multi-protein complexes, Crumbs (Crb)–PALS1 (Stardust)–PATJ (DiscsLost)1,2,3,4 and Cdc42–Par6–Par3–atypical protein kinase C (aPKC), have been implicated in the assembly of tight junctions and in polarization of Drosophila melanogaster epithelia5,6,7,8. Here we identify a biochemical and functional link between these two complexes that is mediated by Par6 and PALS1 (proteins associated with Lin7). The interaction between Par6 and PALS1 is direct, requires the amino terminus of PALS1 and the PDZ domain of Par6, and is regulated by Cdc42-GTP. The transmembrane protein Crb can recruit wild-type Par6, but not Par6 with a mutated PDZ domain, to the cell surface. Expression of dominant-negative PALS1-associated tight junction protein (PATJ) in MDCK cells results in mis-localization of PALS1, members of the Par3–Par6–aPKC complex and the tight junction marker, ZO-1. Similarly, overexpression of Par6 in MDCK cells inhibits localization of PALS1 to the tight junction. Our data highlight a previously unrecognized link between protein complexes that are essential for epithelial polarity and formation of tight junctions.

Access through your institution
Buy or subscribe

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

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

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

Figure 1: Par6 binds PALS1.
Figure 2: Characterization of Par6–PALS1 interaction.
Figure 3: CRB3 recruits Par6 to the plasma membrane.
Figure 4: PATJ(1–238) disrupts aPKCζ localization and tight junction biogenesis.

Similar content being viewed by others

References

  1. Bhat, M.A. et al. Discs Lost, a novel multi-PDZ domain protein, establishes and maintains epithelial polarity. Cell 96, 833–845 (1999).

    Article  CAS  Google Scholar 

  2. Hong, Y., Stronach, B., Perrimon, N., Jan, L.Y. & Jan, Y.N. Drosophila Stardust interacts with Crumbs to control polarity of epithelia but not neuroblasts. Nature 414, 634–638 (2001).

    Article  CAS  Google Scholar 

  3. Bachmann, A., Schneider, M., Thellenberg, E., Grawe, F. & Knust, E. Drosophila Stardust is a partner of Crumbs in the control of epithelial cell polarity. Nature 414, 638–643 (2001).

    Article  CAS  Google Scholar 

  4. Roh, M.H. et al. The Maguk protein, Pals1, functions as an adapter, linking mammalian homologues of Crumbs and Discs Lost. J. Cell Biol. 157, 161–172 (2002).

    Article  CAS  Google Scholar 

  5. Wodarz, A., Ramrath, A., Grimm, A. & Knust, E. Drosophila atypical protein kinase C associates with Bazooka and controls polarity of epithelia and neuroblasts. J. Cell Biol. 150, 1361–1374 (2000).

    Article  CAS  Google Scholar 

  6. Petronczki, M. & Knoblich, J.A. DmPAR-6 directs epithelial polarity and asymmetric cell division of neuroblasts in Drosophila. Nature Cell Biol. 3, 43–49 (2001).

    Article  CAS  Google Scholar 

  7. Joberty, G., Petersen, C., Gao, L. & Macara, I.G. The cell-polarity protein Par6 links Par3 and atypical protein kinase C to Cdc42. Nature Cell Biol. 2, 531–539 (2000).

    Article  CAS  Google Scholar 

  8. Lin, D. et al. A mammalian PAR-3–PAR-6 complex implicated in Cdc42/Rac1 and aPKC signalling and cell polarity. Nature Cell Biol. 2, 540–547 (2000).

    Article  CAS  Google Scholar 

  9. Yeaman, C., Grindstaff, K.K. & Nelson, W.J. New perspectives on mechanisms involved in generating epithelial cell polarity. Physiol. Rev. 79, 73–98 (1999).

    Article  CAS  Google Scholar 

  10. Hung, A.Y. & Sheng, M. PDZ domains: structural modules for protein complex assembly. J. Biol. Chem. 277, 5699–5702 (2002).

    Article  CAS  Google Scholar 

  11. Wodarz, A. Establishing cell polarity in development. Nature Cell Biol. 4, E39–E44 (2002).

    Article  CAS  Google Scholar 

  12. Tepass, U. & Knust, E. Crumbs and stardust act in a genetic pathway that controls the organization of epithelia in Drosophila melanogaster. Dev. Biol. 159, 311–326 (1993).

    Article  CAS  Google Scholar 

  13. Doerks, T. et al. L27, a novel heterodimerization domain in receptor targeting proteins Lin-2 and Lin-7. Trends Biochem. Sci. 25, 317–318 (2000).

    Article  CAS  Google Scholar 

  14. Tepass, U. Adherens junctions: new insight into assembly, modulation and function. Bioessays 24, 690–695 (2002).

    Article  CAS  Google Scholar 

  15. Gao, L., Joberty, G. & Macara, I.G. Assembly of epithelial tight junctions is negatively regulated by Par6. Curr. Biol. 12, 221–225 (2002).

    Article  CAS  Google Scholar 

  16. Suzuki, A. et al. Atypical protein kinase C is involved in the evolutionarily conserved par protein complex and plays a critical role in establishing epithelia-specific junctional structures. J. Cell Biol. 152, 1183–1196 (2001).

    Article  CAS  Google Scholar 

  17. Qiu, R.G., Abo, A. & Martin, S.G. A human homolog of the C. elegans polarity determinant Par-6 links Rac and Cdc42 to PKCζ signaling and cell transformation. Curr. Biol. 10, 697–707 (2000).

    Article  CAS  Google Scholar 

  18. Tabuse, Y. et al. Atypical protein kinase C cooperates with PAR-3 to establish embryonic polarity in Caenorhabditis elegans. Development 125, 3607–3614 (1998).

    CAS  PubMed  Google Scholar 

  19. Izumi, Y. et al. An atypical PKC directly associates and colocalizes at the epithelial tight junction with ASIP, a mammalian homologue of Caenorhabditis elegans polarity protein PAR-3. J. Cell Biol. 143, 95–106 (1998).

    Article  CAS  Google Scholar 

  20. Makarova, O., Roh, M.H., Liu, C.J., Laurinec, S. & Margolis, B. Mammalian Crumbs3 is small transmembrane protein linked to Pals1. Gene (in the press).

  21. Lemmers, C. et al. hINADl/PATJ, a homolog of Discs lost, interacts with crumbs and localizes to tight junctions in human epithelial cells. J. Biol. Chem. 277, 25408–25415 (2002).

    Article  CAS  Google Scholar 

  22. Itoh, M. et al. Junctional adhesion molecule (JAM) binds to PAR-3: a possible mechanism for the recruitment of PAR-3 to tight junctions. J. Cell Biol. 154, 491–497 (2001).

    Article  CAS  Google Scholar 

  23. Ebnet, K. et al. The cell polarity protein ASIP/PAR-3 directly associates with junctional adhesion molecule (JAM). EMBO J. 20, 3738–3748 (2001).

    Article  CAS  Google Scholar 

  24. 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).

    Article  CAS  Google Scholar 

  25. Horne-Badovinac, S. et al. Positional cloning of heart and soul reveals multiple roles for PKC λ in zebrafish organogenesis. Curr. Biol. 11, 1492–1502 (2001).

    Article  CAS  Google Scholar 

  26. Etienne-Manneville, S. & Hall, A. Integrin-mediated activation of Cdc42 controls cell polarity in migrating astrocytes through PKCζ. Cell 106, 489–498 (2001).

    Article  CAS  Google Scholar 

  27. Gotta, M., Abraham, M.C. & Ahringer, J. CDC-42 controls early cell polarity and spindle orientation in C. elegans. Curr. Biol. 11, 482–488 (2001).

    Article  CAS  Google Scholar 

  28. Kay, A.J. & Hunter, C.P. CDC-42 regulates PAR protein localization and function to control cellular and embryonic polarity in C. elegans. Curr. Biol. 11, 474–481 (2001).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank S. Laurinec for antibody generation and A. Liu for assistance with tissue culture and confocal microscopy. We thank A. Saltiel for helpful discussions. We acknowledge the University of Michigan Microscopy and Image Analysis Core for allowing us to use the confocal microscope. This work was partially supported by grant CA40042 from the National Institutes of Health, DHHS (to I.G.M). B.M. is an investigator of the Howard Hughes Medical Institute.

Author information

Author notes

  1. Toby W. Hurd and Lin Gao: These authors contributed equally to this work

Authors and Affiliations

  1. Howard Hughes Medical Institute, University of Michigan Medical School, Ann Arbor, 48109-0650, MI, USA

    Toby W. Hurd & Ben Margolis

  2. Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, 48109-0650, MI, USA

    Michael H. Roh & Ben Margolis

  3. Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, 48109-0650, MI, USA

    Ben Margolis

  4. Center for Cell Signalling, University of Virginia Health Sciences Center, Charlottesville, 22908-0577, VA, USA

    Lin Gao & Ian G. Macara

Authors
  1. Toby W. Hurd
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Lin Gao
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Michael H. Roh
    View author publications

    You can also search for this author inPubMed Google Scholar

  4. Ian G. Macara
    View author publications

    You can also search for this author inPubMed Google Scholar

  5. Ben Margolis
    View author publications

    You can also search for this author inPubMed Google Scholar

Corresponding authors

Correspondence to Ian G. Macara or Ben Margolis.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hurd, T., Gao, L., Roh, M. et al. Direct interaction of two polarity complexes implicated in epithelial tight junction assembly. Nat Cell Biol 5, 137–142 (2003). https://doi.org/10.1038/ncb923

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb923

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