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

Abstract

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.

A Continuous Bovine Kidney Cell Line Constitutively Expressing Bovine αVβ6Integrin Has Increased Susceptibility to Foot-and-Mouth Disease Virus

“…LFBK cells (18) were propagated in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum and antibiotics as described previously (19) and were used for these experiments between passages 64 and 71. LFBK-␣ V ␤ 6 cells were propagated in DMEM (catalog no.…”

Calpain-dependent cytoskeletal rearrangement exploited for anthrax toxin endocytosis

“…We used regulated transcription from a lentivirusbased human EST library to perturb host gene expression globally and randomly (14,15,36,37) in murine RAW267.4 macrophages and isolated 96 macrophage colonies that survived exposure to the PA-LF complex under conditions that normally are lethal. Doxycycline, which represses a modified CMV pro-…”

Upregulation of the Host SLC11A1 Gene by Clostridium difficile Toxin B Facilitates Glucosylation of Rho GTPases and Enhances Toxin Lethality

“…Whereas some host genes defend against detrimental effects of infections, others are exploited to assist pathogen entry, propagation, or release from infected cells or to promote the actions of toxic moieties produced by the pathogen (18)(19)(20)(21)(22)(23). Pathogen-exploited host genes have been termed CGEPs (for cellular genes exploited by pathogens) (22,24).…”

“…Whereas some host genes defend against detrimental effects of infections, others are exploited to assist pathogen entry, propagation, or release from infected cells or to promote the actions of toxic moieties produced by the pathogen (18)(19)(20)(21)(22)(23). Pathogen-exploited host genes have been termed CGEPs (for cellular genes exploited by pathogens) (22,24). Earlier work from our laboratory and others has established the usefulness of random gene inactivation and phenotype-based genetic screening to identify such genes (20,22,(25)(26)(27).…”

“…Pathogen-exploited host genes have been termed CGEPs (for cellular genes exploited by pathogens) (22,24). Earlier work from our laboratory and others has established the usefulness of random gene inactivation and phenotype-based genetic screening to identify such genes (20,22,(25)(26)(27).…”