Baculovirus Lymantria dispar multiple nucleopolyhedrovirus IAP2 and IAP3 do not suppress apoptosis, but trigger apoptosis of insect cells in a transient expression assay

Hayato Yamada, Miyuki Shibuya, Michihiro Kobayashi, Motoko Ikeda

Ld652Y cells derived from the gypsy moth, Lymantria dispar, are permissive for productive infection with L. dispar multiple nucleopolyhedrovirus (LdMNPV), but undergo apoptosis upon infection with various other NPVs, including those isolated from Bombyx mori, Hyphantria cunea, Spodoptera exigua, Orgyia pseudotsugata, and Spodoptera litura. In this study, we examined whether LdMNPV-encoded inhibitor of apoptosis 2 (Ld-IAP2) and 3 (Ld-IAP3) are involved in apoptosis suppression in LdMNPV-infected Ld652Y cells. We found that neither Ld-IAP2 nor Ld-IAP3 was able to suppress the apoptosis of Ld652Y cells induced by p35-defective Autographa californica MNPV (vAcΔp35). However, both Ld-IAP2 and Ld-IAP3 induced apoptosis in Ld652Y cells in a transient expression assay. The apoptosis induced by Ld-IAP3 was accompanied by the stimulation of caspase-3-like protease activity and cleavage of the B. mori homolog of the initiator caspase Dronc, and was precluded by the LdMNPV-encoded apoptosis suppressor protein Apsup and H. cunea MNPV IAP3. Inconsistent with the results obtained previously in SpIm, Ld652Y and High Five cells infected with NPVs from H. cunea, O. pseudotsugata, and A. californica, respectively, considerable stimulation of caspase-3-like protease activity was not observed in LdMNPV-infected Ld652Y cells, likely due to the strong apoptosis suppression activity of Apsup. These results, together with the previous finding that RNAi-mediated silencing of apsup induces apoptosis of LdMNPV-infected Ld652Y cells, indicate that Apsup, but not Ld-IAP2 or Ld-IAP3, is primarily responsible for the suppression of apoptosis in LdMNPV-infected Ld652Y cells. However, it remains inconclusive whether Ld-IAP2 and Ld-IAP3 function as pro-apoptotic proteins in LdMNPV-infected Ld652Y cells.