Genome-scale RNAi on living-cell microarrays identifies novel regulators of Drosophila melanogaster TORC1–S6K pathway signaling

摘要: The evolutionarily conserved target of rapamycin complex 1 (TORC1) controls cell growth in response to nutrient availability and factors. TORC1 signaling is hyperactive cancer, regulators may rep- resent therapeutic targets for human diseases. To identify novel signaling, we performed a genome- scale RNA interference screen on microarrays Drosophila melanogaster cells expressing RPS6, effector whose phosphorylated form detected by immunofluorescence. Our revealed that the TORC1-S6K-RPS6 sig- naling axis regulated many subcellular components, including Class I vesicle coat (COPI), spliceosome, proteasome, nuclear pore, translation initiation machinery. Using additional RNAi reagents Western blotting, confirmed 70 genes as significant on-target RPS6 phosphorylation, characterized them with extensive secondary assays probing various arms pathways, identifying functional relationships among those genes. We conclude cell-based are useful platform genome-scale screening Drosophila, revealing represent drug cancers other diseases deregulated signaling. (Supplemental material available this article.)