The human respiratory syncytial virus nonstructural protein 1 regulates type I and type II interferon pathways

Marcus L. Hastie, Madeleine J. Headlam, Nirav B. Patel, Alexander Bukreyev, Ursula J. Buchholz, Keyur A. Dave, Emma L. Norris, Cassandra L. Wright, Kirsten M. Spann, Peter L. Collins, Jeffrey J. Gorman

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Respiratory syncytial viruses encode a nonstructural protein (NS1) that interferes with type I and III interferon and other antiviral responses. Proteomic studies were conducted on human A549 type II alveolar epithelial cells and type I interferon-deficient Vero cells (African green monkey kidney cells) infected with wild-type and NS1-deficient clones of human respiratory syncytial virus to identify other potential pathway and molecular targets of NS1 interference. These analyses included two-dimensional differential gel electrophoresis and quantitative Western blotting. Surprisingly, NS1 was found to suppress the induction of manganese superoxide dismutase (SOD2) expression in A549 cells and to a much lesser degree Vero cells in response to infection. Because SOD2 is not directly inducible by type I interferons, it served as a marker to probe the impact of NS1 on signaling of other cytokines known to induce SOD2 expression and/or indirect effects of type I interferon signaling. Deductive analysis of results obtained from cell infection and cytokine stimulation studies indicated that interferon-γ signaling was a potential target of NS1, possibly as a result of modulation of STAT1 levels. However, this was not sufficient to explain the magnitude of the impact of NS1 on SOD2 induction in A549 cells. Vero cell infection experiments indicated that NS1 targeted a component of the type I interferon response that does not directly induce SOD2 expression but is required to induce another initiator of SOD2 expression. STAT2 was ruled out as a target of NS1 interference using quantitative Western blot analysis of infected A549 cells, but data were obtained to indicate that STAT1 was one of a number of potential targets of NS1. A label-free mass spectrometry-based quantitative approach is proposed as a means of more definitive identification of NS1 targets.

Original languageEnglish (US)
Pages (from-to)108-127
Number of pages20
JournalMolecular and Cellular Proteomics
Volume11
Issue number5
DOIs
StatePublished - May 2012

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Human respiratory syncytial virus
Interferon Type I
Viruses
Interferon-gamma
Vero Cells
Interferons
Proteins
Cytokines
Infection
Western Blotting
Alveolar Epithelial Cells
Cercopithecus aethiops
Electrophoresis
Respiratory Syncytial Viruses
Superoxide Dismutase
Antiviral Agents
Mass spectrometry
Electrophoresis, Gel, Two-Dimensional
Labels
Gels

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Cite this

The human respiratory syncytial virus nonstructural protein 1 regulates type I and type II interferon pathways. / Hastie, Marcus L.; Headlam, Madeleine J.; Patel, Nirav B.; Bukreyev, Alexander; Buchholz, Ursula J.; Dave, Keyur A.; Norris, Emma L.; Wright, Cassandra L.; Spann, Kirsten M.; Collins, Peter L.; Gorman, Jeffrey J.

In: Molecular and Cellular Proteomics, Vol. 11, No. 5, 05.2012, p. 108-127.

Research output: Contribution to journalArticle

Hastie, ML, Headlam, MJ, Patel, NB, Bukreyev, A, Buchholz, UJ, Dave, KA, Norris, EL, Wright, CL, Spann, KM, Collins, PL & Gorman, JJ 2012, 'The human respiratory syncytial virus nonstructural protein 1 regulates type I and type II interferon pathways', Molecular and Cellular Proteomics, vol. 11, no. 5, pp. 108-127. https://doi.org/10.1074/mcp.M111.015909
Hastie, Marcus L. ; Headlam, Madeleine J. ; Patel, Nirav B. ; Bukreyev, Alexander ; Buchholz, Ursula J. ; Dave, Keyur A. ; Norris, Emma L. ; Wright, Cassandra L. ; Spann, Kirsten M. ; Collins, Peter L. ; Gorman, Jeffrey J. / The human respiratory syncytial virus nonstructural protein 1 regulates type I and type II interferon pathways. In: Molecular and Cellular Proteomics. 2012 ; Vol. 11, No. 5. pp. 108-127.
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