RT Journal Article
SR Electronic
T1 A549 cell expression of ubiquitin E3 ligase RNF168 is decreased following RSV infection but increased by interferon beta treatment
JF European Respiratory Journal
JO Eur Respir J
FD European Respiratory Society
SP P1206
VO 42
IS Suppl 57
A1 Jonathan Jeyatheswaran
A1 Louise Apperley
A1 Paul McNamara
A1 Brian Flanagan
YR 2013
UL http://erj.ersjournals.com/content/42/Suppl_57/P1206.abstract
AB IntroductionRespiratory syncitial virus (RSV) is a major cause of lower respiratory tract illness in young children worldwide. We have shown previously that infection with RSV alters expression of protein linked to intrinsic antiviral defense within the epithelium, the site of initial RSV infection. SiRNA knockdown of the E3 ubiquitin ligase RNF168 in A549 cells leads to increased replication of RSV, suggesting this protein limits viral replication. RNF168 has been shown to be degraded during Herpes Simplex virus infection.AimsTo examine the interaction between RSV and RNF168 and determine if RSV infection leads to reduced RNF168 expression.MethodA549 cells were infected with RSV strain A2 at multiplicity of infection (MOI) up to 1:1. Samples were collected at 4 and 24 hours post infection and RNF168 expression measured by western blot. To examine the mechanism of down regulation, comparison was made to treatment with interferon and the TLR3 stimulant polyI:C.ResultsMaximal change in expression was observed at 4 and 24 hours. An MOI of 1:1 resulted in a three-fold decrease in RNF168 expression compared to control at both 4 hour and 24 hours (n=3). PolyI:C and interferon both increased RNF168 expression by three-fold at 4 hours compared to control.ConclusionRSV infection results in reduced expression of RNF168, suggesting this protein is important in cellular defense against RSV. This suggestion is supported by the observed increase in RNF168 expression following interferon stimulation.