The Ca2+-dependent protease Calpain A regulates Cactus/IκB levels during Drosophila development in response to maternal Dpp signals

Regulation of NFκB activity is central to many processes during development and disease. Activation of NFκB family members depends on degradation of inhibitory IκB proteins. In Drosophila, a nuclear gradient of the NFκB/c-rel protein Dorsal subdivides the embryonic dorsal–ventral axis, defining the extent and location of mesodermal and ectodermal territories. Activation of the Toll pathway directs Dorsal nuclear translocation by inducing proteosomal degradation of the IκB homologue Cactus. Another mechanism that impacts on Dorsal activation involves the Toll-independent pathway, which regulates constitutive Cactus degradation. We have shown that the BMP protein Decapentaplegic (Dpp) inhibits Cactus degradation independent of Toll. Here we report on a novel element of this pathway: the calcium-dependent protease Calpain A. CalpainA knockdowns increase Cactus levels, shifting the Dorsal gradient and dorsal–ventral patterning. As shown for mammalian IκB, this effect requires PEST sequences in the Cactus C-terminus, implying a conserved role for calpains. Alteration of CalpainA or dpp results in similar effects on Dorsal target genes. Epistatic analysis confirms CalpainA activity is regulated by Dpp, indicating that Dpp signals increase Cactus levels through Calpain A inhibition, thereby interfering with Dorsal activation. This mechanism may allow coordination of Toll, BMP and Ca2+ signals, conferring precision to Dorsal-target expression domains.