Activation pathways and signal-mediated upregulation of the insect Spodoptera frugiperda caspase-1.

Sf-caspase-1 is the most studied effector caspase of Lepidoptera. Its activation is believed to follow a two-step mechanism: The first step requires cleavage by an initiator caspase at D195 (between the large and small subunits) releasing the C-terminal small subunit. This is blocked by the baculovirus caspase inhibitor P49. The second step removes the N-terminal prodomain by cleavage at D28 to generate the large subunit that is blocked by the baculovirus caspase inhibitor P35. In this study, we identified an alternative mechanism of Sf-caspase-1 activation. This additional two-step mechanism involves first cleavage of pro-Sf-caspase-1 at D28 to remove the N-terminal prodomain and subsequently cleavage at D195 to generate the large and small subunits. Both mechanisms are triggered by apoptotic stimuli following a distinct pattern. We also showed that expression of Sf-caspase-1 was upregulated upon reception of apoptotic stimuli. Different from all published data, this upregulation occurred as a post-transcriptional event. Moreover, we proved that the stronger the stimuli, the higher the upregulation. And we demonstrated that P49 and P35 inhibited the cleavage at D28 and D195 respectively, independently of wether the first cleavage was at D195 or at D28.