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Published online 9 August 2004. doi:10.1083/jcb.200311031
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 166, Number 4, 549-557
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Article

DaPKC-dependent phosphorylation of Crumbs is required for epithelial cell polarity in Drosophila



Sol Sotillos, María Teresa Díaz-Meco, Eva Caminero, Jorge Moscat, and Sonsoles Campuzano

Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas and Universidad Autonóma de Madrid, Cantoblanco, 28049 Madrid, Spain

Address correspondence to Sonsoles Campuzano, Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Cientificas and Univerisidad Autonoma de Madrid, Cantoblanco, 28049 Madrid, Spain. Tel.: 34-91-497-5072. Fax: 34-91-497-4799. email: scampuzano{at}cbm.uam.es

Both in Drosophila and vertebrate epithelial cells, the establishment of apicobasal polarity requires the apically localized, membrane-associated Par-3–Par-6–aPKC protein complex. In Drosophila, this complex colocalizes with the Crumbs–Stardust (Sdt)–Pals1-associated TJ protein (Patj) complex. Genetic and molecular analyses suggest a functional relationship between them. We show, by overexpression of a kinase-dead Drosophila atypical PKC (DaPKC), the requirement for the kinase activity of DaPKC to maintain the position of apical determinants and to restrict the localization of basolateral ones. We demonstrate a novel physical interaction between the apical complexes, via direct binding of DaPKC to both Crb and Patj, and identify Crumbs as a phosphorylation target of DaPKC. This phosphorylation of Crumbs is functionally significant. Thus, a nonphosphorylatable Crumbs protein behaves in vivo as a dominant negative. Moreover, the phenotypic effect of overexpressing wild-type Crumbs is suppressed by reducing DaPKC activity. These results provide a mechanistic framework for the functional interaction between the Par-3–Par-6–aPKC and Crumbs–Sdt–Patj complexes based in the posttranslational modification of Crb by DaPKC.

Key Words: DaPKC; Crumbs; Drosophila; epithelial cell polarity; signal transduction

Abbreviations used in this paper: ASIP, atypical PKC isotype-specific interacting protein; Baz, Bazooka; Crbi, intracellular domain of Crumbs; DaPKC, Drosophila atypical PKC; Dlg, Discs large; DN, dominant negative; Lgl, Lethal giant larvae; Nrt, Neurotactin; Patj, Pals1-associated TJ protein; Scrib, Scribble; Sdt, Stardust; SP, signal peptide; TJ, tight junctions; TM, transmembrane; ZA, zonula adherens.


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