Genetic proteomic and functional analysis of junctional complexes in Drosophila

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Our bodies rely on the integrity of our barrier epithelia; the sheets of cells (like skin, gut, lung) that separate us from the outside world, or keep different regions of our bodies apart. The thousands of cells in such epithelia have highly specialised junctions that zip them together, in order to prevent leakage (a 'tight' epithelium) or to permit flux of only specific solutes (a 'leaky' epithelium). Problems with this integrity can be serious or fatal (e.g. peritonitis, multiple sclerosis, kidney disease). What makes a junction either tight or leaky? Many of the proteins that are found in junctions are already known. However, with the exceptions of some human genetic diseases and a few 'knockout' mouse lines, it is hard to tease apart the contributions of each protein. We propose to study the septate junction (the homologue of the vertebrate 'tight' junction) in the kidney of the model insect, Drosophila melanogaster. This is an ideal system, because we can cheaply and quickly intervene in gene function in cells of our choice. We will use proteomics to identify the major proteins in the junction, then mutate the genes implicated by the proteomics results, and study whether the junctions remain tight, or become leaky. This will allow a comprehensive study of the relative roles of the proteins that make up the junction.