Role of peroxiredoxin 1 and peroxiredoxin 4 in protection of respiratory syncytial virus-induced cysteinyl oxidation of nuclear cytoskeletal proteins

Mohammad Jamaluddin, John E. Wiktorowicz, Kizhake V. Soman, Istvan Boldogh, Jeffrey D. Forbus, Heidi Spratt, Roberto Garofalo, Allan R. Brasier

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

The respiratory epithelium plays a central role in innate immunity by secreting networks of inflammatory mediators in response to respiratory syncytial virus (RSV) infection. Previous proteomic studies focusing on the host cellular response to RSV indicated the existence of a nuclear heat shock response and cytoplasmic depletion of antioxidant proteins in model type II-like airway epithelial cells. Here, we increased the depth of nuclear proteomic interrogation by using fluorescence difference labeling followed by liquid isoelectric focusing prefractionation/two-dimensional gel electrophoresis (2-DE) to identify an additional 41 proteins affected by RSV infection. Surprisingly, we found inducible oligomers and shifts in isoelectric points for peroxiredoxin 1 (Prdx-1), Prdx-3, and Prdx-4 isoforms without changes in their total abundance, indicating that Prdxs were being oxidized in response to RSV. To address the role of Prdx-1 and Prdx-4 in RSV infection, isoforms were selectively knocked down by small interfering RNA (siRNA) transfection. Cells lacking Prdx-1, Prdx-4, or both showed increased levels of reactive oxygen species formation and a higher level of protein carbonylation in response to RSV infection. Using a novel saturation fluorescence labeling 2-DE analysis, we showed that 15 unique proteins had enhanced oxidative modifications of at least >1.2-fold in the Prdx knockdowns in response to RSV, including annexin A2 and desmoplakin. Our results suggest that Prdx-1 and Prdx-4 are essential for preventing RSV-induced oxidative damage in a subset of nuclear intermediate filament and actin binding proteins in epithelial cells.

Original languageEnglish (US)
Pages (from-to)9533-9545
Number of pages13
JournalJournal of Virology
Volume84
Issue number18
DOIs
StatePublished - 2010

Fingerprint

peroxiredoxin
Peroxiredoxins
Respiratory Syncytial Virus Infections
cytoskeletal proteins
Cytoskeletal Proteins
Respiratory Syncytial Viruses
nuclear proteins
Nuclear Proteins
oxidation
viruses
Proteomics
Protein Isoforms
Desmoplakins
Fluorescence
Epithelial Cells
Protein Carbonylation
Annexin A2
Heat-Shock Response
Microfilament Proteins
Respiratory Mucosa

ASJC Scopus subject areas

  • Immunology
  • Virology
  • Medicine(all)

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Role of peroxiredoxin 1 and peroxiredoxin 4 in protection of respiratory syncytial virus-induced cysteinyl oxidation of nuclear cytoskeletal proteins. / Jamaluddin, Mohammad; Wiktorowicz, John E.; Soman, Kizhake V.; Boldogh, Istvan; Forbus, Jeffrey D.; Spratt, Heidi; Garofalo, Roberto; Brasier, Allan R.

In: Journal of Virology, Vol. 84, No. 18, 2010, p. 9533-9545.

Research output: Contribution to journalArticle

Jamaluddin, Mohammad ; Wiktorowicz, John E. ; Soman, Kizhake V. ; Boldogh, Istvan ; Forbus, Jeffrey D. ; Spratt, Heidi ; Garofalo, Roberto ; Brasier, Allan R. / Role of peroxiredoxin 1 and peroxiredoxin 4 in protection of respiratory syncytial virus-induced cysteinyl oxidation of nuclear cytoskeletal proteins. In: Journal of Virology. 2010 ; Vol. 84, No. 18. pp. 9533-9545.
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