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
References
Andrejeva J, Young DF, Goodbourn S, Randall RE (2002) Degradation of STAT1 and STAT2 by the V proteins of simian virus 5 and human parainfluenza virus type 2, respectively: consequences for virus replication in the presence of alpha/beta and gamma interferons. J Virol 76(5):2159–2167
Baum A, Garcia-Sastre A (2011) Differential recognition of viral RNA by RIG-I. Virulence 2(2):166–169
Beattie E, Denzler KL, Tartaglia J, Perkus ME, Paoletti E, Jacobs BL (1995) Reversal of the interferon-sensitive phenotype of a vaccinia virus lacking E3L by expression of the reovirus S4 gene. J Virol 69(1):499–505
Bellido T, O’Brien CA, Roberson PK, Manolagas SC (1998) Transcriptional activation of the p21(WAF1, CIP1, SDI1) gene by interleukin-6 type cytokines. A prerequisite for their pro-differentiating and anti-apoptotic effects on human osteoblastic cells. J Biol Chem 273(33):21137–21144
Caraglia M, Marra M, Pelaia G, Maselli R, Caputi M, Marsico SA, Abbruzzese A (2005) Alpha-interferon and its effects on signal transduction pathways. J Cell Physiol 202(2):323–335. doi:10.1002/jcp.20137
Catlett-Falcone R, Landowski TH, Oshiro MM, Turkson J, Levitzki A, Savino R, Ciliberto G, Moscinski L, Fernandez-Luna JL, Nunez G, Dalton WS, Jove R (1999) Constitutive activation of Stat3 signaling confers resistance to apoptosis in human U266 myeloma cells. Immunity 10(1):105–115
Chakrabarti A, Jha BK, Silverman RH (2011) New insights into the role of RNase L in innate immunity. J Interf Cytokine Res Off J Int Soc Interferon Cytokine Res 31(1):49–57
Chandesris MO, Melki I, Natividad A, Puel A, Fieschi C, Yun L, Thumerelle C, Oksenhendler E, Boutboul D, Thomas C, Hoarau C, Lebranchu Y, Stephan JL, Cazorla C, Aladjidi N, Micheau M, Tron F, Baruchel A, Barlogis V, Palenzuela G, Mathey C, Dominique S, Body G, Munzer M, Fouyssac F, Jaussaud R, Bader-Meunier B, Mahlaoui N, Blanche S, Debre M, Le Bourgeois M, Gandemer V, Lambert N, Grandin V, Ndaga S, Jacques C, Harre C, Forveille M, Alyanakian MA, Durandy A, Bodemer C, Suarez F, Hermine O, Lortholary O, Casanova JL, Fischer A, Picard C (2012) Autosomal dominant STAT3 deficiency and hyper-IgE syndrome: molecular, cellular, and clinical features from a French national survey. Medicine (Baltimore) 91(4):e1–e19. doi:10.1097/MD.0b013e31825f95b9
Chappell VL, Le LX, LaGrone L, Mileski WJ (2000) Stat proteins play a role in tumor necrosis factor alpha gene expression. Shock 14(3):400–402
Choi HJ, Han JS (2012) Overexpression of phospholipase D enhances Bcl-2 expression by activating STAT3 through independent activation of ERK and p38MAPK in HeLa cells. Biochim Biophys Acta 1823(6):1082–1091. doi:10.1016/j.bbamcr.2012.03.015
Clemens MJ, Elia A (1997) The double-stranded RNA-dependent protein kinase PKR: structure and function. J Interferon Cytokine Res 17(9):503–524
Collins AS, Ahmed S, Napoletano S, Schroeder M, Johnston JA, Hegarty JE, O’Farrelly C, Stevenson NJ (2014) Hepatitis C virus (HCV)-induced suppressor of cytokine signaling (SOCS) 3 regulates proinflammatory TNF-alpha responses. J Leukoc Biol 96(2):255–263. doi:10.1189/jlb.2A1211-608RRRR
Dai J, Pan W, Wang P (2011) ISG15 facilitates cellular antiviral response to dengue and west nile virus infection in vitro. Virol J 8:468. doi:10.1186/1743-422x-8-468
Darnell JE Jr, Kerr IM, Stark GR (1994) Jak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins. Science 264(5164):1415–1421
Elliott J, Lynch O, Suessmuth Y, Qian P, Boyd C, Burrows J, Buick R, Stevenson N, Touzelet O, Gadina M, Power U, Johnston J (2007) Respiratory syncytial virus NS1 protein degrades STAT2 by using the Elongin-Cullin E3 ligase. J Virol 81(7):3428–3436 (JVI.02303-06)
Epperson DE, Arnold D, Spies T, Cresswell P, Pober JS, Johnson DR (1992) Cytokines increase transporter in antigen processing-1 expression more rapidly than HLA class I expression in endothelial cells. J Immunol 149(10):3297–3301
Galabru J, Hovanessian A (1987) Autophosphorylation of the protein kinase dependent on double-stranded RNA. J Biol Chem 262(32):15538–15544
Garcia R, Yu CL, Hudnall A, Catlett R, Nelson KL, Smithgall T, Fujita DJ, Ethier SP, Jove R (1997) Constitutive activation of Stat3 in fibroblasts transformed by diverse oncoproteins and in breast carcinoma cells. Cell Growth Differ 8(12):1267–1276
Garcin D, Marq JB, Strahle L, le Mercier P, Kolakofsky D (2002) All four Sendai Virus C proteins bind Stat1, but only the larger forms also induce its mono-ubiquitination and degradation. Virology 295(2):256–265
Gouilleux-Gruart V, Debierre-Grockiego F, Gouilleux F, Capiod JC, Claisse JF, Delobel J, Prin L (1997) Activated Stat related transcription factors in acute leukemia. Leuk Lymphoma 28(1–2):83–88
Goujon C, Moncorge O, Bauby H, Doyle T, Ward CC, Schaller T, Hue S, Barclay WS, Schulz R, Malim MH (2013) Human MX2 is an interferon-induced post-entry inhibitor of HIV-1 infection. Nature 502(7472):559–562
Gritsko T, Williams A, Turkson J, Kaneko S, Bowman T, Huang M, Nam S, Eweis I, Diaz N, Sullivan D, Yoder S, Enkemann S, Eschrich S, Lee JH, Beam CA, Cheng J, Minton S, Muro-Cacho CA, Jove R (2006) Persistent activation of stat3 signaling induces survivin gene expression and confers resistance to apoptosis in human breast cancer cells. Clin Cancer Res 12(1):11–19
Guerra S, Caceres A, Knobeloch KP, Horak I, Esteban M (2008) Vaccinia virus E3 protein prevents the antiviral action of ISG15. PLoS Pathog 4(7):e1000096. doi:10.1371/journal.ppat.1000096
Haller O, Staeheli P, Schwemmle M, Kochs G (2015) Mx GTPases: dynamin-like antiviral machines of innate immunity. Trends Microbiol 23(3):154–163. doi:10.1016/j.tim.2014.12.003
Hilbert DM, Kopf M, Mock BA, Kohler G, Rudikoff S (1995) Interleukin 6 is essential for in vivo development of B lineage neoplasms. J Exp Med 182(1):243–248
Ho HH, Ivashkiv LB (2006) Role of STAT3 in type I interferon responses. Negative regulation of STAT1-dependent inflammatory gene activation. J Biol Chem 281(20):14111–14118
Holland SM, DeLeo FR, Elloumi HZ, Hsu AP, Uzel G, Brodsky N, Freeman AF, Demidowich A, Davis J, Turner ML, Anderson VL, Darnell DN, Welch PA, Kuhns DB, Frucht DM, Malech HL, Gallin JI, Kobayashi SD, Whitney AR, Voyich JM, Musser JM, Woellner C, Schaffer AA, Puck JM, Grimbacher B (2007) STAT3 mutations in the hyper-IgE syndrome. N Engl J Med 357(16):1608–1619
Honda K, Yanai H, Takaoka A, Taniguchi T (2005) Regulation of the type I IFN induction: a current view. Int Immunol 17(11):1367–1378. doi:10.1093/intimm/dxh318
Horvath CM, Stark GR, Kerr IM, Darnell JE (1996) Interactions between STAT and non-STAT proteins in the interferon-stimulated gene factor 3 transcription complex. Mol Cell Biol 16(12):6957–6964
Huang M, Page C, Reynolds RK, Lin J (2000) Constitutive activation of stat 3 oncogene product in human ovarian carcinoma cells. Gynecol Oncol 79(1):67–73
Isaacs A, Lindenmann J (1957) Virus interference. I. The interferon. Proc R Soc Lond B Biol Sci 147(927):258–267
Kinjyo I, Inoue H, Hamano S, Fukuyama S, Yoshimura T, Koga K, Takaki H, Himeno K, Takaesu G, Kobayashi T, Yoshimura A (2006) Loss of SOCS3 in T helper cells resulted in reduced immune responses and hyperproduction of interleukin 10 and transforming growth factor-beta 1. J Exp Med 203(4):1021–1031. doi:10.1084/jem.20052333
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227(5259):680–685
Le Bon A, Schiavoni G, D’Agostino G, Gresser I, Belardelli F, Tough DF (2001) Type i interferons potently enhance humoral immunity and can promote isotype switching by stimulating dendritic cells in vivo. Immunity 14(4):461–470
Le Bon A, Thompson C, Kamphuis E, Durand V, Rossmann C, Kalinke U, Tough DF (2006) Cutting edge: enhancement of antibody responses through direct stimulation of B and T cells by type I IFN. J Immunol 176(4):2074–2078
Lou W, Ni Z, Dyer K, Tweardy DJ, Gao AC (2000) Interleukin-6 induces prostate cancer cell growth accompanied by activation of stat3 signaling pathway. Prostate 42(3):239–242
Mali SB (2015) Review of STAT3 (signal transducers and activators of transcription) in head and neck cancer. Oral Oncol 51(6):565–569. doi:10.1016/j.oraloncology.2015.03.004
Myskiw C, Arsenio J, van Bruggen R, Deschambault Y, Cao J (2009) Vaccinia virus E3 suppresses expression of diverse cytokines through inhibition of the PKR, NF-kappaB, and IRF3 pathways. J Virol 83(13):6757–6768
Nielsen M, Kaltoft K, Nordahl M, Ropke C, Geisler C, Mustelin T, Dobson P, Svejgaard A, Odum N (1997) Constitutive activation of a slowly migrating isoform of Stat3 in mycosis fungoides: tyrphostin AG490 inhibits Stat3 activation and growth of mycosis fungoides tumor cell lines. Proc Natl Acad Sci USA 94(13):6764–6769
Niu G, Briggs J, Deng J, Ma Y, Lee H, Kortylewski M, Kujawski M, Kay H, Cress WD, Jove R, Yu H (2008) Signal transducer and activator of transcription 3 is required for hypoxia-inducible factor-1alpha RNA expression in both tumor cells and tumor-associated myeloid cells. Mol Cancer Res 6(7):1099–1105
Perdiguero B, Esteban M (2009) The interferon system and vaccinia virus evasion mechanisms. J Interferon Cytokine Res Off J Int Soc Interferon Cytokine Res 29(9):581–598
Qureshi SA, Salditt-Georgieff M, Darnell JE Jr (1995) Tyrosine-phosphorylated Stat1 and Stat2 plus a 48-kDa protein all contact DNA in forming interferon-stimulated-gene factor 3. Proc Natl Acad Sci USA 92(9):3829–3833
Salazar-Mather TP, Ishikawa R, Biron CA (1996) NK cell trafficking and cytokine expression in splenic compartments after IFN induction and viral infection. J Immunol 157(7):3054–3064
Shuai K (1994) Interferon-activated signal transduction to the nucleus. Curr Opin Cell Biol 6(2):253–259
Siegel AM, Heimall J, Freeman AF, Hsu AP, Brittain E, Brenchley JM, Douek DC, Fahle GH, Cohen JI, Holland SM, Milner JD (2011) A critical role for STAT3 transcription factor signaling in the development and maintenance of human T cell memory. Immunity 35(5):806–818
Stark GR, Kerr IM, Williams BR, Silverman RH, Schreiber RD (1998) How cells respond to interferons. Annu Rev Biochem 67:227–264. doi:10.1146/annurev.biochem.67.1.227
Stevenson NJ, Bourke NM, Ryan EJ, Binder M, Fanning L, Johnston JA, Hegarty JE, Long A, O’Farrelly C (2013) Hepatitis C virus targets the interferon-alpha JAK/STAT pathway by promoting proteasomal degradation in immune cells and hepatocytes. FEBS Lett 587(10):1571–1578
Ulane CM, Rodriguez JJ, Parisien JP, Horvath CM (2003) STAT3 ubiquitylation and degradation by mumps virus suppress cytokine and oncogene signaling. J Virol 77(11):6385–6393
Unterholzner L (2013) The interferon response to intracellular DNA: why so many receptors? Immunobiology 218(11):1312–1321. doi:10.1016/j.imbio.2013.07.007
Wang T, Niu G, Kortylewski M, Burdelya L, Shain K, Zhang S, Bhattacharya R, Gabrilovich D, Heller R, Coppola D, Dalton W, Jove R, Pardoll D, Yu H (2004) Regulation of the innate and adaptive immune responses by Stat-3 signaling in tumor cells. Nat Med 10(1):48–54
Wang WB, Levy DE, Lee CK (2011) STAT3 negatively regulates type I IFN-mediated antiviral response. J Immunol 187(5):2578–2585
Yang E, Lerner L, Besser D, Darnell JE Jr (2003) Independent and cooperative activation of chromosomal c-fos promoter by STAT3. J Biol Chem 278(18):15794–15799
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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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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DOI: https://doi.org/10.1007/s00018-016-2435-3