Insect cuticular melanization is regulated by the prophenoloxidase (proPO)-activating system, which is a component of innate immunity. However, the differentiation between cuticular melanization and innate immunity is not well defined. Here, we demonstrate that the proPO-activating system regulates cuticular melanization in the silkworm pupae using a different mechanism. Our results indicate that the differential and spatial regulation of key components, such as the proPO-activating factor, tyrosine hydroxylase, proPOs, and immulectin, primes the proPO-activating system for either cuticular melanization or innate immunity. This dual strategy for cuticular melanization in development and innate immunity upon infection demonstrates a two-pronged defense mechanism mediated by the priming of the proPO system.