Vesicular stomatitis viruses expressing wild-type or mutant M proteins activate apoptosis through distinct pathways.
Vesicular stomatitis virus (VSV) induces apoptosis by at least two mechanisms. The viral matrix (M) protein induces apoptosis via the mitochondrial pathway due to the inhibition of host gene expression. However, in some cell types, the inhibition of host gene expression by VSV expressing wild-type (wt) M protein delays VSV-induced apoptosis, indicating that another mechanism is involved. In support of this, the recombinant M51R-M (rM51R-M) virus, expressing a mutant M protein that is defective in its ability to inhibit host gene expression, induces apoptosis much more rapidly in L929 cells than do viruses expressing wt M protein. Here, we determine the caspase pathways by which the rM51R-M virus induces apoptosis. An analysis of caspase activity, using fluorometric caspase assays and Western blots, indicated that each of the main initiator caspases, caspase-8, caspase-9, and caspase-12, were activated during infection with the rM51R-M virus. The overexpression of Bcl-2, an inhibitor of the mitochondrial pathway, or MAGE-3, an inhibitor of caspase-12 activation, did not delay apoptosis induction in rM51R-M virus-infected L929 cells. However, an inhibitor of caspase-8 activity significantly delayed apoptosis induction. Furthermore, the inhibition of caspase-8 activity prevented the activation of caspase-9, suggesting that caspase-9 is activated by cross talk with caspase-8. These data indicate that VSV expressing the mutant M protein induces apoptosis via the death receptor apoptotic pathway, a mechanism distinct from that induced by VSV expressing the wt M protein.
Gaddy
Daniel F
DF
Department of Microbiology and Immunology, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA. dgaddy@wfubmc.edu
Lyles
Douglas S
DS
D019943:Amino Acid Substitution; D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D053454:Caspase 12; D053181:Caspase 8; D053453:Caspase 9; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D004789:Enzyme Activation; D005470:Fluorometry; D051379:Mice; D009154:Mutation; D009363:Neoplasm Proteins / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000502:physiology; D014721:Vesicular stomatitis Indiana virus / Q000502:physiology*; D014763:Viral Matrix Proteins / Q000235:genetics* / Q000502:physiology*
D061945:Caspase Inhibitors; C094460:M protein, Vesicular stomatitis virus; D009363:Neoplasm Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D014763:Viral Matrix Proteins; C506440:Casp12 protein, mouse; C505526:Casp8 protein, mouse; C506434:Casp9 protein, mouse; D053454:Caspase 12; D053181:Caspase 8; D053453:Caspase 9; D020169:Caspases