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

Abstract：

: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.

journal_name

Virology

journal_title

Virology

authors

Kolokoltsova OA,Domina AM,Kolokoltsov AA,Davey RA,Weaver SC,Watowich SJ

doi

10.1016/j.virol.2008.03.025

subject

Has Abstract

pub_date

2008-07-20 00:00:00

pages

133-42

issue

1

eissn

0042-6822

issn

1096-0341

pii

S0042-6822(08)00195-5

journal_volume

377

pub_type

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