The polarized architecture of epithelial cells depends on the highly stereotypic distribution of cellular junctions and other membrane-associated protein complexes. In epithelial cells of the Drosophila embryo, three distinct domains subdivide the lateral plasma membrane. The most apical one comprises the subapical complex (SAC). It is followed by the zonula adherens (ZA) and, further basally, by the septate junction. A core component of the SAC is the transmembrane protein Crumbs, the cytoplasmic domain of which recruits the PDZ-protein Discs Lost into the complex. Cells lacking crumbs or the functionally related gene stardust fail to organize a continuous ZA and to maintain cell polarity. Here we show that stardust provides an essential component of the SAC. Stardust proteins colocalize with Crumbs and bind to the carboxy-terminal amino acids of its cytoplasmic tail. We introduce two different Stardust proteins here: one MAGUK protein, characterized by a PDZ domain, an SH3 domain and a guanylate kinase domain; and a second isoform comprising only the guanylate kinase domain. The Stardust proteins represent versatile candidates as structural and possibly regulatory constituents of the SAC, a crucial element in the control of epithelial cell polarity.
Significance
We report the first to our knowledge genetically engineered honeybees, which are important pollinators and interesting biological models for the study of social and complex behaviors as well as caste and sexual development. This genetic manipulation tool will enable systematic studies of biological processes in an organism building complex societies. We demonstrate highly efficient integration and expression of
piggyBac
-derived cassettes in the honeybee that make this system applicable to colony-based screening approaches and useful for an average beekeeping facility. This cassette was stably and efficiently transmitted and expressed in progeny by two different promoters, offering the prospect for activation or inhibition of gene functions under conditions of stage- and tissue-specific promoters.
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