Respiratory syncytial virus infection induces a reactive oxygen species-MSK1-phospho-Ser-276 RelA pathway required for cytokine expression.
Respiratory syncytial virus (RSV) is a human pathogen that induces airway inflammation, at least in part, by modulating gene expression programs in airway epithelial cells. The presence of RSV replication is detected by the intracellular retinoic acid-inducible gene I (RIG-I) RNA helicase that forms a productive signaling complex with the mitochondrion-anchored MAVS protein, resulting in nuclear translocation of the NF-kappaB transcription factor. Although nuclear translocation is a prerequisite for activation of the innate inflammatory response, recent studies show that separate pathways governing RelA activation are also required for target gene expression. In this study, we examine the mechanism of RelA phosphorylation and its requirement for RSV-induced gene expression. RSV infection produced a time-dependent RelA phosphorylation on serine (Ser) residues Ser-276 and Ser-536 in parallel with enhanced reactive oxygen species (ROS) stress. Inhibition of RSV-induced ROS inhibited formation of phospho-Ser-276 RelA without affecting phospho-Ser-536 RelA formation. RSV potently induced activation of cytoplasmic mitogen- and stress-related kinase 1 (MSK1) in an ROS-dependent manner. Inhibition of MSK1 using H89 and small interfering RNA knockdown both reduced RSV-induced phospho-Ser-276 RelA formation and expression of a subset of NF-kappaB-dependent genes. Direct examination of the role of phospho-Ser-276 in target gene expression by expression of a RelA Ser-276-to-Ala site mutation in RelA(-/-) mouse embryonic fibroblasts showed that the mutation was unable to mediate RSV-induced NF-kappaB-dependent gene expression. We conclude that RSV induces RelA activation in the innate inflammatory response via a pathway separate from that controlling RelA cytoplasmic release, mediated by ROS signaling to cytoplasmic MSK1 activation and RelA Ser-276 phosphorylation.
Jamaluddin
Mohammad
M
Institute for Translational Sciences, University of Texas Medical Branch (UTMB), Galveston, TX 77555-1060, USA.
Tian
Bing
B
Boldogh
Istvan
I
Garofalo
Roberto P
RP
Brasier
Allan R
AR
D000818:Animals; D016207:Cytokines / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology / Q000821:virology; D005347:Fibroblasts / Q000166:cytology / Q000821:virology; D005786:Gene Expression Regulation; D006801:Humans; D008168:Lung / Q000166:cytology / Q000821:virology; D051379:Mice; D010766:Phosphorylation; D017382:Reactive Oxygen Species / Q000378:metabolism*; D018113:Respiratory Syncytial Virus, Human / Q000472:pathogenicity*; D038744:Ribosomal Protein S6 Kinases, 90-kDa / Q000235:genetics / Q000378:metabolism*; D012694:Serine / Q000378:metabolism; D051996:Transcription Factor RelA / Q000378:metabolism*
D016207:Cytokines; D017382:Reactive Oxygen Species; D051996:Transcription Factor RelA; D012694:Serine; D038744:Ribosomal Protein S6 Kinases, 90-kDa; C116919:mitogen and stress-activated protein kinase 1
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