ABSTRACT
Caspases provide vital links in non-apoptotic regulatory networks controlling inflammation, compensatory proliferation, morphology and cell migration. How caspases are activated under non-apoptotic conditions and process a selective set of substrates without killing the cell remain enigmatic. Here we find that the Drosophila unconventional myosin CRINKLED (CK) selectively interacts with the initiator caspase DRONC and regulates some of its non-apoptotic functions. Loss of CK in the arista, border cells or proneural clusters of the wing imaginal discs affects DRONC-dependent patterning. Our data indicate that CK acts as substrate adaptor, recruiting SHAGGY46/GSK3-ß to DRONC, thereby facilitating caspase-mediated cleavage and localized modulation of kinase activity. Similarly, the mammalian CK counterpart, MYO7A, binds to and impinges on CASPASE-8, revealing a new regulatory axis affecting receptor interacting protein kinase-1 (RIPK1)>CASPASE-8 signalling. Together, our results expose a conserved role for unconventional myosins in transducing caspase-dependent regulation of kinases, allowing them to take part in specific signalling events.
Subject(s)
Caspase 8/metabolism , Caspases/metabolism , Drosophila Proteins/metabolism , Myosins/metabolism , Animals , Cell Line, Tumor , Drosophila melanogaster , Flow Cytometry , Glycogen Synthase Kinase 3/metabolism , Glycogen Synthase Kinase 3 beta , Humans , Immunoprecipitation , Mice , Microscopy, Confocal , Myosin VIIa , NIH 3T3 Cells , Receptor-Interacting Protein Serine-Threonine Kinases/metabolism , Signal Transduction , Wings, AnimalABSTRACT
Class I phosphoinositide 3-kinases (PI(3)Ks) are activated through associated adaptor molecules in response to G protein-coupled and tyrosine kinase receptor signalling. They contain Ras-binding domains (RBDs) and can also be activated through direct association with active GTP-bound Ras. The ability of Ras to activate PI(3)K has been established in vitro and by overexpression analysis, but its relevance for normal PI(3)K function in vivo is unknown. The Drosophila class I PI(3)K, Dp110, is activated by nutrient-responsive insulin signalling and modulates growth, oogenesis and metabolism. To investigate the importance of Ras-mediated PI(3)K activation for normal PI(3)K function, we replaced Dp110 with Dp110(RBD), which is unable to bind to Ras but otherwise biochemically normal. We found that Ras-mediated Dp110 regulation is dispensable for viability. However, egg production, which requires large amounts of growth, is dramatically lowered in Dp110(RBD) flies. Furthermore, insulin cannot maximally activate PI(3)K signalling in Dp110(RBD) imaginal discs and Dp110(RBD) flies are small. Thus, Dp110 integrates inputs from its phosphotyrosine-binding adaptor and Ras to achieve maximal PI(3)K signalling in specific biological situations.
Subject(s)
Drosophila melanogaster/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Signal Transduction , ras Proteins/metabolism , Animals , Binding Sites/genetics , Brain/drug effects , Brain/growth & development , Brain/metabolism , Cell Survival , Drosophila Proteins/genetics , Drosophila Proteins/metabolism , Drosophila melanogaster/cytology , Drosophila melanogaster/genetics , Enzyme Activation/drug effects , Female , Immunoblotting , Insulin/pharmacology , Male , Microscopy, Fluorescence , Mutation/genetics , Phosphatidylinositol 3-Kinases/genetics , Phosphorylation/drug effects , Protein Binding , Wings, Animal/drug effects , Wings, Animal/growth & development , Wings, Animal/metabolism , ras Proteins/geneticsABSTRACT
In the fruit fly Drosophila melanogaster, the insulin and ecdysone signaling pathways have long been known to regulate growth and developmental timing, respectively. Recent findings reveal that crosstalk between these pathways allows coordination of growth and developmental timing and thus determines final body size.
