Alphavirus production is inhibited in neurofibromin 1-deficient cells through activated RAS signalling

Journal Article · Sun Jul 20 04:00:00 EDT 2008 · Virology

DOI:https://doi.org/10.1016/j.virol.2008.03.025· OSTI ID:21141013

Davey, Robert A ^[1]; Weaver, Scott C ^[1]; Watowich, Stanley J ^[2]

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1. Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX (United States)
2. Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX (United States)

Virus-host interactions essential for alphavirus pathogenesis are poorly understood. To address this shortcoming, we coupled retrovirus insertional mutagenesis and a cell survival selection strategy to generate clonal cell lines broadly resistant to Sindbis virus (SINV) and other alphaviruses. Resistant cells had significantly impaired SINV production relative to wild-type (WT) cells, although virus binding and fusion events were similar in both sets of cells. Analysis of the retroviral integration sites identified the neurofibromin 1 (NF1) gene as disrupted in alphavirus-resistant cell lines. Subsequent analysis indicated that expression of NF1 was significantly reduced in alphavirus-resistant cells. Importantly, independent down-regulation of NF1 expression in WT HEK 293 cells decreased virus production and increased cell viability during SINV infection, relative to infected WT cells. Additionally, we observed hyperactive RAS signalling in the resistant HEK 293 cells, which was anticipated because NF1 is a negative regulator of RAS. Expression of constitutively active RAS (HRAS-G12V) in a WT HEK 293 cell line resulted in a marked delay in virus production, compared with infected cells transfected with parental plasmid or dominant-negative RAS (HRAS-S17N). This work highlights novel host cell determinants required for alphavirus pathogenesis and suggests that RAS signalling may play an important role in neuronal susceptibility to SINV infection.

• OSTI ID: 21141013
• Journal Information: Virology, Journal Name: Virology Journal Issue: 1 Vol. 377; ISSN VIRLAX; ISSN 0042-6822
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
GENE REGULATION
GENES
HOST
INTERACTIONS
MUTAGENESIS
PATHOGENESIS
VIRUSES