ARS | Publication request: Phenotype-Based identification of host genes required for propagation of African swine fever virus

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: October 25, 2006
Publication Date: December 4, 2006
Citation: Zsak, L., Chang, A.Y., Feng, Y., Mosseri, R., Lu, Q., Kowalski, P., Zsak, A., Burrage, T.G., Neilan, J.G., Kutish, G.F., Lu, Z., Laegreid, W.W., Rock, D.L., Cohen, S.N. 2006. Phenotype-Based identification of host genes required for propagation of African swine fever virus [abstract]. In: Proceedings of the 87th Conference of Research Workers in Animal Diseases, December 3-5, 2006, Chicago, Illinois. p. 128.

Technical Abstract: African Swine Fever Virus (ASFV) produces a fatal acute hemorrhagic fever in domesticated pigs that is of worldwide economic importance. Using an expressed sequence tag (EST)-library-based antisense method of random gene inactivation and a phenotypic screen for limitation of ASFV propagation in cultured human cells, we identified six host genes whose cellular functions are exploited by ASFV. These included three loci, BAT3 (HLA-B associated transcript 3), C1qTNF (C1q and tumor necrosis factor related protein 6), and TOM40 (translocase of outer mitochondrial membrane 40), whose antisense expression from a tetracycline (Tet)-regulated promoter resulted in reversible inhibition of ASFV production by >99%. A BAT3 EST that encodes amino acid residues 450 to 518 of the mature protein and also limited ASFV production reversibly when expressed in the sense orientiation was investigated more extensively. Sense expression of the BAT3 peptide, which appears to functionally inactivate the coexisting chromosomal BAT3 gene by a dominant negative mechanism, resulted in decreased synthesis of viral DNA and proteins early after ASFV infection, altered expression of apoptosis-related genes as determined by cDNA microarray analysis, and increased cellular sensitivity to staurosporin-induced apoptosis. Antisense transcription of BAT3 reduced ASFV production by a mechanism that did not detectably affect the synthesis of virus macromolecules. Our results, which demonstrate the utility of EST-based functional screens for the detection of host genes exploited by pathogenic viruses, reveal a novel collection of cellular genes previously not known to be required for ASFV infection.