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Journal of Virology, September 2006, p. 8705-8717, Vol. 80, No. 17
0022-538X/06/$08.00+0     doi:10.1128/JVI.00475-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Phenotype-Based Identification of Host Genes Required for Replication of African Swine Fever Virus{dagger}

Annie C. Y. Chang,1,{ddagger} Laszlo Zsak,2,{ddagger} Yanan Feng,1 Ronen Mosseri,1,§ Quan Lu,1 Paul Kowalski,1 Aniko Zsak,2 Thomas G. Burrage,2 John G. Neilan,2 Gerald F. Kutish,2 Zhiqiang Lu,2 Will Laegreid,3 Daniel L. Rock,2,¶ and Stanley N. Cohen1,4*

Departments of Genetics,1 Medicine, Stanford University School of Medicine, Stanford, California,4 Plum Island Animal Disease Center, Orient Point, New York,2 Meat Animal Research Center, Lincoln, Nebraska3

Received 7 March 2006/ Accepted 2 June 2006

African swine fever virus (ASFV) produces a fatal acute hemorrhagic fever in domesticated pigs that potentially is a worldwide economic threat. Using an expressed sequence tag (EST) library-based antisense method of random gene inactivation and a phenotypic screen for limitation of ASFV replication in cultured human cells, we identified six host genes whose cellular functions are required 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), for which antisense expression from a tetracycline-regulated promoter resulted in reversible inhibition of ASFV production by >99%. The effects of antisense transcription of the BAT3 EST and also of expression in the sense orientation of this EST, which encodes amino acid residues 450 to 518 of the mature BAT3 protein, were investigated more extensively. Sense expression of the BAT3 peptide, which appears to reversibly interfere with BAT3 function by a dominant negative mechanism, resulted in decreased synthesis of viral DNA and proteins early after ASFV infection, altered transcription of apoptosis-related genes as determined by cDNA microarray analysis, and increased cellular sensitivity to staurosporine-induced apoptosis. Antisense transcription of BAT3 reduced ASFV production without affecting abundance of the virus macromolecules we assayed. 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.

* Corresponding author. Mailing address: Stanford University School of Medicine, Department of Genetics, 300 Pasteur Drive, Stanford, CA 94305-5120. Phone: (650) 723-5315. Fax: (750) 725-1536. E-mail: sncohen{at}stanford.edu.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

{ddagger} A.C.Y.C. and L.Z. contributed equally to the work.

§ Present address: Kahanman Pediatric Health Clinic, Leumit HMO, Bnei-Brak, Israel.

Present address: Department of Pathobiology, College of Veterinary Medicine, University of Illinois at Urbana-Champaign, Urbana, Ill.

Journal of Virology, September 2006, p. 8705-8717, Vol. 80, No. 17
0022-538X/06/$08.00+0     doi:10.1128/JVI.00475-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Piccone, M. E., Feng, Y., Chang, A. C. Y., Mosseri, R., Lu, Q., Kutish, G. F., Lu, Z., Burrage, T. G., Gooch, C., Rock, D. L., Cohen, S. N. (2009). Identification of Cellular Genes Affecting the Infectivity of Foot-and-Mouth Disease Virus. J. Virol. 83: 6681-6688 [Abstract] [Full Text]  


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