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

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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.

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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

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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.

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Authors and Affiliations

  1. School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute (TBSI), Trinity College Dublin, Dublin, Ireland

    Rebecca Mahony, Siobhán Gargan, Sinead E. Keating, Andrew G. Bowie, Cliona O’Farrelly & Nigel J. Stevenson

  2. Department of Microbiology, Moyne Institute of Preventive Medicine, School of Genetics and Microbiology, Trinity College Dublin, Dublin, Ireland

    Kim L. Roberts

  3. School of Medicine, Trinity Biomedical Sciences Institute (TBSI), Trinity College Dublin, Dublin, Ireland

    Cliona O’Farrelly

  4. Centre for Innate Immunity and Infectious Diseases, Hudson Institute of Medical Research, Clayton, VIC, Australia

    Nollaig Bourke

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  1. Rebecca Mahony
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  4. Nollaig Bourke
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Correspondence to Nigel J. Stevenson.

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Mahony, R., Gargan, S., Roberts, K.L. et al. A novel anti-viral role for STAT3 in IFN-α signalling responses. Cell. Mol. Life Sci. 74, 1755–1764 (2017). https://doi.org/10.1007/s00018-016-2435-3

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