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Journal of Virology, July 2009, p. 6681-6688, Vol. 83, No. 13
0022-538X/09/$08.00+0     doi:10.1128/JVI.01729-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Identification of Cellular Genes Affecting the Infectivity of Foot-and-Mouth Disease Virus{triangledown}

Maria E. Piccone,1,§ Yanan Feng,2,§ Annie C. Y. Chang,2 Ronen Mosseri,2 Quan Lu,2,¶ Gerald F. Kutish,1 Zhiqiang Lu,1 Thomas G. Burrage,1 Christina Gooch,1,{dagger} Daniel L. Rock,1,{ddagger} and Stanley N. Cohen2,3*

Plum Island Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Greenport, New York 11944,1 Departments of Genetics,2 Medicine, Stanford University School of Medicine, Stanford, California 943053

Received 14 August 2008/ Accepted 3 April 2009

Foot-and-mouth disease virus (FMDV) produces one of the most infectious of all livestock diseases, causing extensive economic loss in areas of breakout. Like other viral pathogens, FMDV recruits proteins encoded by host cell genes to accomplish the entry, replication, and release of infectious viral particles. To identify such host-encoded proteins, we employed an antisense RNA strategy and a lentivirus-based library containing approximately 40,000 human expressed sequence tags (ESTs) to randomly inactivate chromosomal genes in a bovine kidney cell line (LF-BK) that is highly susceptible to FMDV infection and then isolated clones that survived multiple rounds of exposure to the virus. Here, we report the identification of ESTs whose expression in antisense orientation limited host cell killing by FMDV and restricted viral propagation. The role of one such EST, that of ectonucleoside triphosphate diphosphohydrolase 6 (NTPDase6; also known as CD39L2), a membrane-associated ectonucleoside triphosphate diphosphohydrolase that previously was not suspected of involvement in the propagation of viral pathogens and which we now show is required for normal synthesis of FMDV RNA and proteins, is described in this report.

* 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

{triangledown} Published ahead of print on 15 April 2009.

§ These authors contributed equally to the work.

Present address: Department of Environmental Health, Harvard School of Public Health, Boston, MA 02115.

{dagger} Present address: Fred Hutchinson Cancer Research Center, Seattle, WA 98109.

{ddagger} Present address: Department of Pathobiology, College of Veterinary Medicine, University of Illinois at Urbana-Champaign, Urbana, IL 61802.

Journal of Virology, July 2009, p. 6681-6688, Vol. 83, No. 13
0022-538X/09/$08.00+0     doi:10.1128/JVI.01729-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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