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Cellular and Molecular Life Sciences

, Volume 74, Issue 9, pp 1755–1764 | Cite as

A novel anti-viral role for STAT3 in IFN-α signalling responses

  • Rebecca Mahony
  • Siobhán Gargan
  • Kim L. Roberts
  • Nollaig Bourke
  • Sinead E. Keating
  • Andrew G. Bowie
  • Cliona O’Farrelly
  • Nigel J. StevensonEmail author
Original Article

Abstract

The cytokine, Interferon (IFN)-α, induces a wide spectrum of anti-viral mediators, via the Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway. STAT1 and STAT2 are well characterised to upregulate IFN-stimulated gene (ISG) expression; but even though STAT3 is also activated by IFN-α, its role in anti-viral ISG induction is unclear. Several viruses, including Hepatitis C and Mumps, reduce cellular STAT3 protein levels, via the promotion of ubiquitin-mediated proteasomal degradation. This viral immune evasion mechanism suggests an undiscovered anti-viral role for STAT3 in IFN-α signalling. To investigate STAT3’s functional involvement in this Type I IFN pathway, we first analysed its effect upon the replication of two viruses, Influenza and Vaccinia. Viral plaque assays, using Wild Type (WT) and STAT3-/- Murine Embryonic Fibroblasts (MEFs), revealed that STAT3 is required for the inhibition of Influenza and Vaccinia replication. Furthermore, STAT3 shRNA knockdown also enhanced Influenza replication and hindered induction of several, well characterised, anti-viral ISGs: PKR, OAS2, MxB and ISG15; while STAT3 expression had no effect upon induction of a separate ISG group: Viperin, IFI27, CXCL10 and CCL5. These discoveries reveal, for the first time, an anti-viral role for STAT3 in the IFN-α pathway and characterise a requirement for STAT3 in the expression of specific ISGs. These findings also identify STAT3 as a therapeutic target against viral infection and highlight it as an essential pathway component for endogenous and therapeutic IFN-α responsiveness.

Keywords

Janus kinase/signal transducer of activator of transcription (JAK/STAT) Interferon-alpha (IFN-α) STAT3 IFN-stimulated gene (ISG) Vaccinia virus Influenza virus Anti-viral 

Abbreviations

AA

Amino acid

ANOVA

Analysis of variance

BCL6

B-cell lymphoma 6

DC

Dendritic cell

EIF2A

Eukaryotic translation initiation factor 2A

EMCV

Encephalomyocarditis virus

EMSA

Electrophoretic mobility shift assay

GAS

Gamma-activated sequence

HCV

Hepatitis C virus

HIF-1α

Hypoxia-inducible factor 1 alpha

HIV

Human immunodeficiency virus

IAV

Influenza A virus

IFN

Interferon

IFN-α

Interferon alpha

IFN-αR

IFN alpha receptor

IL

Interleukin

IRF

IFN regulatory factor

ISG

IFN-Stimulated Gene

ISGF3

IFN-stimulated gene factor 3

ISRE

IFN-stimulated response element

JAK

Janus kinase

MDCK

Madin–Darby canine kidney

MEF

Murine embryonic fibroblast

MHC

Major histocompatibility complex

MMP

Matrix metalloproteinase

MxA/MxB

Myxovirus resistance gene A/B

NK

Natural killer

OAS2

2′-5′-Oligoadenylate synthetase 2

PKR

Protein kinase R

PRR

Pathogen recognition receptor

qRT-PCR

Quantitative real time polymerase chain reaction

RIG-I

Retinoic acid-inducible gene I

RK13

Rabbit kidney 13

RPS15

Ribosomal protein S15

SH2

Src Homology 2

shRNA

Short hairpin RNA

SIE

Sis-inducible element

SOCS

Suppressor of cytokine signalling

STAT

Signal transducer and activator of transcription

TNF-α

Tumour necrosis factor alpha

TGF-β

Transforming growth factor beta

Tyk2

Tyrosine kinase 2

VACV

Vaccinia virus

VSV

Vesicular stomatitis virus

WT

Wild type

Notes

Acknowledgements

We would like to thank Prof. Valeria Poli (University of Turin) for MEFs and Prof. Bertrand Jacobs (Arizona State University) for the Vaccinia virus. In addition, we wish to thank the Health Research Board (POR-20120-57) and Science Foundation Ireland (12/IA/1667) for funding this work.

Supplementary material

18_2016_2435_MOESM1_ESM.docx (112 kb)
Supplementary material 1 (DOCX 112 kb)

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Copyright information

© Springer International Publishing 2016

Authors and Affiliations

  1. 1.School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute (TBSI)Trinity College DublinDublinIreland
  2. 2.Department of Microbiology, Moyne Institute of Preventive Medicine, School of Genetics and MicrobiologyTrinity College DublinDublinIreland
  3. 3.School of Medicine, Trinity Biomedical Sciences Institute (TBSI)Trinity College DublinDublinIreland
  4. 4.Centre for Innate Immunity and Infectious DiseasesHudson Institute of Medical ResearchClaytonAustralia

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