Autographa californica M Nucleopolyhedrovirus ProV-CATH is Activated during Infected Cell Death

• DOI: 10.1006/viro.2002.1378
• Publication Date: 2002-10-06T18:49:14Z
• Authors: L.G. Hom; T. Ohkawa; D. Trudeau; L.E. Volkman
• Keywords: AcMNPV V-CATH activation; Enzyme Precursors; 0303 health sciences; Apoptosis; Moths; Spodoptera; Cathepsins; Nucleopolyhedroviruses; Cell Line; 3. Good health; Enzyme Activation; Cysteine Endopeptidases; 03 medical and health sciences; Virology; Animals; Lysosomes; baculovirus cathepsin

V-CATH, a cathepsin L-like cysteine protease encoded by the baculovirus Autographa californica M nucleopolyhedrovirus, has been shown to play an essential role in host liquefaction. Similar to cellular cathepsin L, V-CATH is synthesized as an inactive proenzyme and is activated by cleavage of the propeptide. Previous studies indicated that removal of the propeptide was rapid, occurring as soon as the protein could be detected by Western blot, 22 h postinfection. We found, however, that these results reflected artifactual processing of the proenzyme. When the protease inhibitor E-64 was used to prevent this aberration, we found that proV-CATH accumulated in infected cells and activation did not begin until the onset of cell death, at approximately 80 h postinfection. Western blot analysis of fractions of live and dead cells isolated by fluorescence-activated cell sorting revealed that mature V-CATH was found only in dead cells. The regulation of activation of proV-CATH, therefore, was quite different from that of cellular cathepsins. Acridine orange staining revealed that lysosome integrity was lost in dead cells, an occurrence that could lead to the activation of proV-CATH by lysosomal proteases.