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A polarity complex of mPar-6 and atypical PKC binds, phosphorylates and regulates mammalian Lgl

Nature Cell Biology volume 5, pages 301–308 (2003)Cite this article

Abstract

The evolutionarily conserved proteins Par-6, atypical protein kinase C (aPKC), Cdc42 and Par-3 associate to regulate cell polarity and asymmetric cell division, but the downstream targets of this complex are largely unknown. Here we identify direct physiological interactions between mammalian aPKC, murine Par-6C (mPar-6C) and Mlgl, the mammalian orthologue of the Drosophila melanogaster tumour suppressor Lethal (2) giant larvae. In cultured cell lines and in mouse brain, aPKC, mPar-6C and Mlgl form a multiprotein complex in which Mlgl is targeted for phosphorylation on conserved serine residues. These phosphorylation sites are important for embryonic fibroblasts to polarize correctly in response to wounding and may regulate the ability of Mlgl to direct protein trafficking. Our data provide a direct physical and regulatory link between proteins of distinct polarity complexes, identify Mlgl as a functional substrate for aPKC in cell polarization and indicate that aPKC is directed to cell polarity substrates through a network of protein–protein interactions.

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Figure 1: Identification of Mlgl as a binding partner for mPar-6C.
Figure 2: Mlgl and mPar-6C interact in cells.
Figure 3: mPar-6 and Mlgl have a similar cellular distribution pattern.
Figure 4: Several regions of mPar-6C associate with Mlgl.
Figure 5: mPar-6C CRIB and PDZ domains bind to the N terminus of Mlgl.
Figure 6: Mlgl, aPKC and mPar-6C form an oligomeric complex.
Figure 7: aPKC binds and phosphorylates Mlgl.
Figure 8: Mlgl phosphorylation sites have a role in polarizing migrating cells.

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Acknowledgements

We thank L. Velazquez, S. Gelkop and I. Ernberg for reagents and technical advice; B. Baskin and B. Cox for technical assistance; I. Macara for HA–mPar-6A and mPar-6B cDNAs; and J. Knoblich for sharing data before publication. P.J.P., J.P.F. and A.D.H. are recipients of Canadian Institutes of Health Research (CIHR) postdoctoral fellowships and K.B. is the recipient of an NSERC industrial postgraduate scholarship with MDS-Sciex. This work was supported by grants from the National Cancer Institute of Canada, the CIHR and the Ontario R&D Challenge Fund and Genome Canada. T.P. is a Distinguished Scientist of the CIHR.

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  1. Pamela J. Plant and James P. Fawcett: These authors contributed equally to this work

Authors and Affiliations

  1. Programme in Molecular Biology and Cancer, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, M5G 1X5, Canada

    Pamela J. Plant, James P. Fawcett, Dan C.C. Lin, Amy D. Holdorf, Kathleen Binns, Sarang Kulkarni & Tony Pawson

  2. Department of Medical Genetics and Microbiology, University of Toronto, 1 Kings College Circle, Toronto, M5S 1A8, Canada

    Dan C.C. Lin, Kathleen Binns & Tony Pawson

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Correspondence to Tony Pawson.

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Plant, P., Fawcett, J., Lin, D. et al. A polarity complex of mPar-6 and atypical PKC binds, phosphorylates and regulates mammalian Lgl. Nat Cell Biol 5, 301–308 (2003). https://doi.org/10.1038/ncb948

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