RelA Ser276 phosphorylation-coupled Lys310 acetylation controls transcriptional elongation of inflammatory cytokines in respiratory syncytial virus infection

Allan R. Brasier, Bing Tian, M. Jamaluddin, Mridul K. Kalita, Roberto Garofalo, Muping Lu

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Abstract

Respiratory syncytial virus (RSV) is a negative-sense single-stranded RNA virus responsible for lower respiratory tract infections (LRTIs) in humans. In experimental models of RSV LRTI, the actions of the nuclear factor κB (NF-κB) transcription factor mediate inflammation and pathology. We have shown that RSV replication induces a mitogen-and-stress-related kinase 1 (MSK-1) pathway that activates NF-κB RelA transcriptional activity by a process involving serine phosphorylation at serine (Ser) residue 276. In this study, we examined the mechanism by which phospho-Ser276 RelA mediates expression of the NF-κB-dependent gene network. RelA-deficient mouse embryonic fibroblasts (MEFs) complemented with the RelA Ser276Ala mutant are deficient in CXCL2/Groβ, KC, and interleukin-6 (IL-6) expression, but NFKBIA/IκBα is preserved. We show that RSV-induced RelA Ser276 phosphorylation is required for acetylation at Lys310, an event required for transcriptional activity and stable association of RelA with the activated positive transcriptional elongation factor (PTEF-b) complex proteins, bromodomain 4 (Brd4), and cyclin-dependent kinase 9 (CDK9). In contrast to gene loading pattern of PTEF-b proteins produced by tumor necrosis factor (TNF) stimulation, RSV induces their initial clearance followed by partial reaccumulation coincident with RelA recruitment. The RSV-induced binding patterns of the CDK9 substrate, phospho-Ser2 RNA polymerase (Pol) II, follows a similar pattern of clearance and downstream gene reaccumulation. The functional role of CDK9 was examined using CDK9 small interfering RNA (siRNA) and CDK inhibitors, where RSV-induced NF-κB-dependent gene expression was significantly inhibited. Finally, although RSV induces a transition from short transcripts to fully spliced mRNA in wild-type RelA (RelA WT)-expressing cells, this transition is not seen in cells expressing RelA Ser276Ala. We conclude that RelA Ser276 phosphorylation mediates RelA acetylation, Brd4/CDK9 association, and activation of downstream inflammatory genes by transcriptional elongation in RSV infection.

Original languageEnglish (US)
Pages (from-to)11752-11769
Number of pages18
JournalJournal of Virology
Volume85
Issue number22
DOIs
StatePublished - Nov 2011

Fingerprint

Respiratory Syncytial Virus Infections
Respiratory Syncytial Viruses
acetylation
Cyclin-Dependent Kinase 9
Acetylation
phosphorylation
cytokines
Phosphorylation
cyclin-dependent kinase
Cytokines
viruses
infection
Respiratory Tract Infections
Transcriptional Elongation Factors
respiratory tract diseases
serine
Serine
negative sense, single-stranded RNA viruses
Genes
Virus Attachment

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

RelA Ser276 phosphorylation-coupled Lys310 acetylation controls transcriptional elongation of inflammatory cytokines in respiratory syncytial virus infection. / Brasier, Allan R.; Tian, Bing; Jamaluddin, M.; Kalita, Mridul K.; Garofalo, Roberto; Lu, Muping.

In: Journal of Virology, Vol. 85, No. 22, 11.2011, p. 11752-11769.

Research output: Contribution to journalArticle

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AU - Brasier, Allan R.

AU - Tian, Bing

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AU - Garofalo, Roberto

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