Isolation of an apoptosis suppressor gene of the Spodoptera littoralis nucleopolyhedrovirus

Spodoptera frugiperda SF9 cells infected with mutants of the Autographa californica nucleopolyhedrovirus (AcMNPV) which lack a functional p35 gene undergo apoptosis, aborting the viral infection. The Spodoptera littoralis nucleopolyhedrovirus (SINPV) was able to suppress apoptosis triggered by vΔP35k/pol⁺, an AcMNPV p35 null mutant. To identify the putative apoptotic suppressor gene of SINPV, overlapping cosmid clones representing the entire SINPV genome were individually cotransfected along with genomic DNA of vΔP35K/pol⁺. Using this complementation assay, we isolated a SINPV DNA fragment that was able to rescue the vΔP35K/pol⁺ infection in SF9 cells. By further subcloning and rescue, we identified a novel SINPV gene, Slp49. The Slp49 sequence predicted a 49-kda polypeptide with about 48.8% identity to the AcMNPV apoptotic suppressor P35. SLP49 displays a potential recognition site, TVTDG, for cleavage by death caspases. Recombinant AcMNPVs deficient in p35 bearing the Slp49 gene did not induce apoptosis and showed successful productive infections in SF9 cells, indicating that Slp49 is a functional homologue of p35. A 1.5-kbp Slp49-specific transcript was identified in SF9 cells infected with SINPV or with vAc496, a vΔP35K/pol⁺-recombinant bearing Slp49. The discovery of Slp49 contributes to the identification of important functional motifs conserved in p35-like apoptotic suppressors and to the future isolation of p35-like genes from other baculoviruses.