Role of individual caspases induced by astrovirus on the processing of its structural protein and its release from the cell through a non-lytic mechanism.

Caspases (Casp) activity has been associated with the intracellular proteolytic processing of the structural protein to yield the mature capsid formed by VP70 and with the cell release of human astrovirus (HAstV). This work describes the role of individual Casp on these events. The activity of initiator (-8, -9) and executioner (-3/7) Casp was clearly detected at 12h post-infection. All these proteases were able to cleave VP90 in an in vitro assay, but this processing was blocked in cells transfected with siRNA against Casp-3, -9, but not against Casp-8. In contrast, virus release, observed in the absence of cell lysis, was more drastically affected by either silencing Casp-3 or in the presence of the inhibitor Ac-DEVD-CHO. Cleavage of VP90 to yield VP70 was mapped at motif TYVD(657). These data indicate that the processing of VP90 and the release of HAstV from the cell are two Casp-related, but apparently independent, events.