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Journal of Virology, February 1999, p. 1278-1285, Vol. 73, No. 2
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Isolation of an Apoptosis Suppressor Gene of the Spodoptera littoralis Nucleopolyhedrovirusdagger

Quansheng Du,1,2 Dana Lehavi,1 Ouriel Faktor,3 Yipeng Qi,2 and Nor Chejanovsky1,*

Entomology Department, Institute of Plant Protection, Agricultural Research Organization, The Volcani Center, Bet Dagan 50250,1 and Department of Entomology, Faculty of Agriculture, The Hebrew University of Jerusalem, Rehovot 76100,3 Israel, and Institute of Virology, Wuhan University, Wuhan 430072, People's Republic of China2

Received 16 June 1998/Accepted 27 October 1998

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 (SlNPV) was able to suppress apoptosis triggered by vDelta P35K/pol+, an AcMNPV p35 null mutant. To identify the putative apoptotic suppressor gene of SlNPV, overlapping cosmid clones representing the entire SlNPV genome were individually cotransfected along with genomic DNA of vDelta P35K/pol+. Using this complementation assay, we isolated a SlNPV DNA fragment that was able to rescue the vDelta P35K/pol+ infection in SF9 cells. By further subcloning and rescue, we identified a novel SlNPV 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 SlNPV or with vAc496, a vDelta P35K/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.

* Corresponding author. Mailing address: Entomology Department, Institute of Plant Protection, The Volcani Center, POB 6, Bet Dagan 50250, Israel. Phone: (972)-3-9683694. Fax: (972)-3-9604180. E-mail: ninar{at}netvision.net.il.

dagger Contribution 525/98 from the Agricultural Research Organization, The Volcani Center, Bet Dagan, Israel.

Journal of Virology, February 1999, p. 1278-1285, Vol. 73, No. 2
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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