Interferon-α (IFN-α) is essential for antiviral immunity, but in the absence of matrix metalloproteinase-12 (MMP-12) or IκBα (encoded by NFKBIA) we show that IFN-α is retained in the cytosol of virus-infected cells and is not secreted. Our findings suggest that activated IκBα mediates the export of IFN-α from virus-infected cells and that the inability of cells in Mmp12−/− but not wild-type mice to express IκBα and thus export IFN-α makes coxsackievirus type B3 infection lethal and renders respiratory syncytial virus more pathogenic. We show here that after macrophage secretion, MMP-12 is transported into virus-infected cells. In HeLa cells MMP-12 is also translocated to the nucleus, where it binds to the NFKBIA promoter, driving transcription. We also identified dual-regulated substrates that are repressed both by MMP-12 binding to the substrate's gene exons and by MMP-12–mediated cleavage of the substrate protein itself. Whereas intracellular MMP-12 mediates NFKBIA transcription, leading to IFN-α secretion and host protection, extracellular MMP-12 cleaves off the IFN-α receptor 2 binding site of systemic IFN-α, preventing an unchecked immune response. Consistent with an unexpected role for MMP-12 in clearing systemic IFN-α, treatment of coxsackievirus type B3–infected wild-type mice with a membrane-impermeable MMP-12 inhibitor elevates systemic IFN-α levels and reduces viral replication in pancreas while sparing intracellular MMP-12. These findings suggest that inhibiting extracellular MMP-12 could be a new avenue for the development of antiviral treatments.
Subscribe to Journal
Get full journal access for 1 year
only $18.75 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Wang, B.X. & Fish, E.N. The yin and yang of viruses and interferons. Trends Immunol. 33, 190–197 (2012).
Brunner, K.T., Hurez, D., Mc, C.R. & Benacerraf, B. Blood clearance of P32-labeled vesicular stomatitis and Newcastle disease viruses by the reticuloendothelial system in mice. J. Immunol. 85, 99–105 (1960).
Shapiro, S.D., Kobayashi, D.K. & Ley, T.J. Cloning and characterization of a unique elastolytic metalloproteinase produced by human alveolar macrophages. J. Biol. Chem. 268, 23824–23829 (1993).
Belvisi, M.G. & Bottomley, K.M. The role of matrix metalloproteinases (MMPs) in the pathophysiology of chronic obstructive pulmonary disease (COPD): a therapeutic role for inhibitors of MMPs? Inflamm. Res. 52, 95–100 (2003).
Liang, J. et al. Macrophage metalloelastase accelerates the progression of atherosclerosis in transgenic rabbits. Circulation 113, 1993–2001 (2006).
Curci, J.A., Liao, S., Huffman, M.D., Shapiro, S.D. & Thompson, R.W. Expression and localization of macrophage elastase (matrix metalloproteinase-12) in abdominal aortic aneurysms. J. Clin. Invest. 102, 1900–1910 (1998).
McQuibban, G.A. et al. Inflammation dampened by gelatinase A cleavage of monocyte chemoattractant protein-3. Science 289, 1202–1206 (2000).
Parks, W.C., Wilson, C.L. & Lopez-Boado, Y.S. Matrix metalloproteinases as modulators of inflammation and innate immunity. Nat. Rev. Immunol. 4, 617–629 (2004).
Morrison, C.J., Butler, G.S., Rodriguez, D. & Overall, C.M. Matrix metalloproteinase proteomics: substrates, targets, and therapy. Curr. Opin. Cell Biol. 21, 645–653 (2009).
Houghton, A.M. et al. Macrophage elastase (matrix metalloproteinase-12) suppresses growth of lung metastases. Cancer Res. 66, 6149–6155 (2006).
Dean, R.A. et al. Macrophage-specific metalloelastase (MMP-12) truncates and inactivates ELR+ CXC chemokines and generates CCL2, -7, -8, and -13 antagonists: potential role of the macrophage in terminating polymorphonuclear leukocyte influx. Blood 112, 3455–3464 (2008).
Houghton, A.M., Hartzell, W.O., Robbins, C.S., Gomis-Ruth, F.X. & Shapiro, S.D. Macrophage elastase kills bacteria within murine macrophages. Nature 460, 637–641 (2009).
Lambert, A.L., Mangum, J.B., DeLorme, M.P. & Everitt, J.I. Ultrafine carbon black particles enhance respiratory syncytial virus–induced airway reactivity, pulmonary inflammation, and chemokine expression. Toxicol. Sci. 72, 339–346 (2003).
Samuel, C.E. Antiviral actions of interferons. Clinical Microbiol. Rev. 14, 778–809 (2001).
Guerrero-Plata, A., Casola, A. & Garofalo, R.P. Human metapneumovirus induces a profile of lung cytokines distinct from that of respiratory syncytial virus. J. Virol. 79, 14992–14997 (2005).
Vallabhapurapu, S. & Karin, M. Regulation and function of NF-κB transcription factors in the immune system. Annu. Rev. Immunol. 27, 693–733 (2009).
Garmaroudi, F.S. et al. Pairwise network mechanisms in the host signaling response to coxsackievirus B3 infection. Proc. Natl. Acad. Sci. USA 107, 17053–17058 (2010).
Shimizu-Hirota, R. et al. MT1-MMP- regulates the PI3Kδ.Mi-2/NuRD–dependent control of macrophage immune function. Genes Dev. 26, 395–413 (2012).
Wang, X. et al. Matrix metalloproteinase-7 and ADAM-12 (a disintegrin and metalloproteinase-12) define a signaling axis in agonist-induced hypertension and cardiac hypertrophy. Circulation 119, 2480–2489 (2009).
Mosser, D.M. & Edwards, J.P. Exploring the full spectrum of macrophage activation. Nat. Rev. Immunol. 8, 958–969 (2008).
Kleifeld, O. et al. Isotopic labeling of terminal amines in complex samples identifies protein N-termini and protease cleavage products. Nat. Biotechnol. 28, 281–288 (2010).
Koyanagi, S., Ohdo, S., Yukawa, E. & Higuchi, S. Chronopharmacological study of interferon-alpha in mice. J. Pharmacol. Exp. Ther. 283, 259–264 (1997).
Butler, G.S. & Overall, C.M. Proteomic identification of multitasking proteins in unexpected locations complicates drug targeting. Nat. Rev. Drug Discov. 8, 935–948 (2009).
Dufour, A. & Overall, C.M. Missing the target: matrix metalloproteinase antitargets in inflammation and cancer. Trends Pharmacol. Sci. 34, 233–242 (2013).
Overall, C.M. & Kleifeld, O. Tumour microenvironment—opinion: validating matrix metalloproteinases as drug targets and anti-targets for cancer therapy. Nat. Rev. Cancer 6, 227–239 (2006).
Devel, L. et al. Development of selective inhibitors and substrate of matrix metalloproteinase-12. J. Biol. Chem. 281, 11152–11160 (2006).
Johnson, J.L. et al. A selective matrix metalloproteinase-12 inhibitor retards atherosclerotic plaque development in apolipoprotein E–knockout mice. Arterioscler. Thromb. Vasc. Biol. 31, 528–535 (2011).
Rizza, P., Moretti, F. & Belardelli, F. Recent advances on the immunomodulatory effects of IFN-α: implications for cancer immunotherapy and autoimmunity. Autoimmunity 43, 204–209 (2010).
Cheung, C. et al. Ablation of matrix metalloproteinase-9 increases severity of viral myocarditis in mice. Circulation 117, 1574–1582 (2008).
Starr, A.E. & Overall, C.M. Chapter 13. Characterizing proteolytic processing of chemokines by mass spectrometry, biochemistry, neo-epitope antibodies and functional assays. Methods Enzymol. 461, 281–307 (2009).
Buroker, N.E., Barboza, J. & Huang, J.Y. The IκBα gene is a peroxisome proliferator–activated receptor cardiac target gene. FEBS J. 276, 3247–3255 (2009).
Kleifeld, O. et al. Identifying and quantifying proteolytic events and the natural N terminome by terminal amine isotopic labeling of substrates. Nat. Protoc. 6, 1578–1611 (2011).
Robertson, G. et al. Genome-wide profiles of STAT1 DNA association using chromatin immunoprecipitation and massively parallel sequencing. Nat. Methods 4, 651–657 (2007).
auf dem Keller, U., Prudova, A., Gioia, M., Butler, G.S. & Overall, C.M. A statistics-based platform for quantitative N-terminome analysis and identification of protease cleavage products. Mol. Cell. Proteomics 9, 912–927 (2010).
Prudova, A., auf dem Keller, U., Butler, G.S. & Overall, C.M. Multiplex N-terminome analysis of MMP-2 and MMP-9 substrate degradomes by iTRAQ-TAILS quantitative proteomics. Mol. Cell. Proteomics 9, 894–911 (2010).
Fahlman, R.P., Chen, W. & Overall, C.M. Absolute proteomic quantification of the activity state of proteases and proteolytic cleavages using proteolytic signature peptides and isobaric tags. J. Proteomics 100, 79–91 (2014).
Pedrioli, P.G. Trans-proteomic pipeline: a pipeline for proteomic analysis. Methods Mol. Biol. 604, 213–238 (2010).
Deutsch, E.W. et al. A guided tour of the Trans-Proteomic Pipeline. Proteomics 10, 1150–1159 (2010).
Keller, A., Nesvizhskii, A.I., Kolker, E. & Aebersold, R. Empirical statistical model to estimate the accuracy of peptide identifications made by MS/MS and database search. Anal. Chem. 74, 5383–5392 (2002).
auf dem Keller, U. & Overall, C.M. An add-on to the Trans-Proteomic Pipeline for the Automated Analysis of TAILS data. Biol. Chem. 393, 1477–1483 (2012).
We thank C. Smits at St. Paul's Hospital for technical assistance and expert advice regarding the humane care of animal models used in this study. We thank T. Buroker at Seattle Children's Hospital for IκBα plasmid promoter constructs and A. Hoffmann at the University of California–San Diego for Nfkbia−/− cells and advice. HL1 cardiomyocytes were a gift from W. Claycomb (Louisiana State University). This work was supported by Canadian Institutes of Health Research grants on MMPs during viral infection (no. 08-0369 (B.M.M., D.J.M.)) and on MMPs in inflammation (nos. MOP-37937 and MOP-111055 (C.M.O.)) and an Infrastructure Grant from the Michael Smith Research Foundation (University of British Columbia Centre for Blood Research) and by the British Columbia Proteomics Network (C.M.O.); salary support for D.J.M. is provided by a Canada Research Chair in Viral Pathogenesis and is supported by research fellowships from the US Myocarditis Foundation and the Heart and Stroke Foundation of Canada; H.L. is funded by the Heart and Stroke Foundation of British Columbia and Yukon; salary support for C.M.O. is provided by a Canada Research Chair in Metalloproteinase Proteomics and Systems Biology B.M.M. is funded by the Heart and Stroke Foundation of British Columbia and Yukon, Genome Canada/British Columbia and the Networks of Centres of Excellence–CECR Centre of Excellence for Prevention of Organ Failure.
The authors declare no competing financial interests.
About this article
Cite this article
Marchant, D., Bellac, C., Moraes, T. et al. A new transcriptional role for matrix metalloproteinase-12 in antiviral immunity. Nat Med 20, 493–502 (2014) doi:10.1038/nm.3508
Vitamin D3-vitamin D receptor axis suppresses pulmonary emphysema by maintaining alveolar macrophage homeostasis and function
Investigative Opthalmology & Visual Science (2019)
The ectoenzyme-side of matrix metalloproteinases (MMPs) makes inflammation by serum amyloid A (SAA) and chemokines go round
Immunology Letters (2019)
British Journal of Pharmacology (2019)