Abstract
Identification of immune effectors and the post-translational modifications that control their activity is essential for dissecting mechanisms of immunity. Here we demonstrate that the antiviral activity of interferon-induced transmembrane protein 3 (IFITM3) is post-translationally regulated by S-palmitoylation. Large-scale profiling of palmitoylated proteins in a dendritic cell line using a chemical reporter strategy revealed over 150 lipid-modified proteins with diverse cellular functions, including innate immunity. We discovered that S-palmitoylation of IFITM3 on membrane-proximal cysteines controls its clustering in membrane compartments and its antiviral activity against influenza virus. The sites of S-palmitoylation are highly conserved among the IFITM family of proteins in vertebrates, which suggests that S-palmitoylation of these immune effectors may be an ancient post-translational modification that is crucial for host resistance to viral infections. The S-palmitoylation and clustering of IFITM3 will be important for elucidating its mechanism of action and for the design of antiviral therapeutics.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Finlay, B.B. & McFadden, G. Anti-immunology: evasion of the host immune system by bacterial and viral pathogens. Cell 124, 767–782 (2006).
Harrison, S.C. Viral membrane fusion. Nat. Struct. Mol. Biol. 15, 690–698 (2008).
Flannagan, R.S., Cosio, G. & Grinstein, S. Antimicrobial mechanisms of phagocytes and bacterial evasion strategies. Nat. Rev. Microbiol. 7, 355–366 (2009).
Linder, M.E. & Deschenes, R.J. Palmitoylation: policing protein stability and traffic. Nat. Rev. Mol. Cell Biol. 8, 74–84 (2007).
Charron, G., Wilson, J. & Hang, H.C. Chemical tools for understanding protein lipidation in eukaryotes. Curr. Opin. Chem. Biol. 13, 382–391 (2009).
Martin, B.R. & Cravatt, B.F. Large-scale profiling of protein palmitoylation in mammalian cells. Nat. Methods 6, 135–138 (2009).
Charron, G. et al. Robust fluorescent detection of protein fatty-acylation with chemical reporters. J. Am. Chem. Soc. 131, 4967–4975 (2009).
Roth, A.F. et al. Global analysis of protein palmitoylation in yeast. Cell 125, 1003–1013 (2006).
Kang, R. et al. Neural palmitoyl-proteomics reveals dynamic synaptic palmitoylation. Nature 456, 904–909 (2008).
Yang, W., Di Vizio, D., Kirchner, M., Steen, H. & Freeman, M.R. Proteome-scale characterization of human S-acylated proteins in lipid raft-enriched and non-raft membranes. Mol. Cell Proteomics 9, 54–70 (2009).
Zhang, J. et al. Identification of CKAP4/p63 as a major substrate of the palmitoyl acyltransferase DHHC2, a putative tumor suppressor, using a novel proteomics method. Mol. Cell. Proteomics 7, 1378–1388 (2008).
Reis e Sousa, C. Dendritic cells in a mature age. Nat. Rev. Immunol. 6, 476–483 (2006).
Steinman, R.M. & Banchereau, J. Taking dendritic cells into medicine. Nature 449, 419–426 (2007).
Shen, Z., Reznikoff, G., Dranoff, G. & Rock, K.L. Cloned dendritic cells can present exogenous antigens on both MHC class I and class II molecules. J. Immunol. 158, 2723–2730 (1997).
Yang, Y.Y., Ascano, J.M. & Hang, H.C. Bioorthogonal chemical reporters for monitoring protein acetylation. J. Am. Chem. Soc. 132, 3640–3641 (2010).
Brass, A.L. et al. The IFITM proteins mediate cellular resistance to influenza A H1N1 virus, West Nile virus, and dengue virus. Cell 139, 1243–1254 (2009).
Saitou, M., Barton, S.C. & Surani, M.A. A molecular programme for the specification of germ cell fate in mice. Nature 418, 293–300 (2002).
Lange, U.C. et al. Normal germ line establishment in mice carrying a deletion of the Ifitm/Fragilis gene family cluster. Mol. Cell. Biol. 28, 4688–4696 (2008).
Smith, R.A., Young, J., Weis, J.J. & Weis, J.H. Expression of the mouse fragilis gene products in immune cells and association with receptor signaling complexes. Genes Immun. 7, 113–121 (2006).
Ni, Z. et al. Apical role for BRG1 in cytokine-induced promoter assembly. Proc. Natl. Acad. Sci. USA 102, 14611–14616 (2005).
Neil, S.J., Zang, T. & Bieniasz, P.D. Tetherin inhibits retrovirus release and is antagonized by HIV-1 Vpu. Nature 451, 425–430 (2008).
Alber, D. & Staeheli, P. Partial inhibition of vesicular stomatitis virus by the interferon-induced human 9–27 protein. J. Interferon Cytokine Res. 16, 375–380 (1996).
Abrami, L., Kunz, B., Iacovache, I. & van der Goot, F.G. Palmitoylation and ubiquitination regulate exit of the Wnt signaling protein LRP6 from the endoplasmic reticulum. Proc. Natl. Acad. Sci. USA 105, 5384–5389 (2008).
Abrami, L., Leppla, S.H. & van der Goot, F.G. Receptor palmitoylation and ubiquitination regulate anthrax toxin endocytosis. J. Cell Biol. 172, 309–320 (2006).
Hemler, M.E. Tetraspanin functions and associated microdomains. Nat. Rev. Mol. Cell Biol. 6, 801–811 (2005).
Schmidt, M.F. & Schlesinger, M.J. Fatty acid binding to vesicular stomatitis virus glycoprotein: a new type of post-translational modification of the viral glycoprotein. Cell 17, 813–819 (1979).
Veit, M., Schmidt, M.F. & Rott, R. Different palmitoylation of paramyxovirus glycoproteins. Virology 168, 173–176 (1989).
Chen, B.J., Takeda, M. & Lamb, R.A. Influenza virus hemagglutinin (H3 subtype) requires palmitoylation of its cytoplasmic tail for assembly: M1 proteins of two subtypes differ in their ability to support assembly. J. Virol. 79, 13673–13684 (2005).
Moltedo, B. et al. Cutting edge: stealth influenza virus replication precedes the initiation of adaptive immunity. J. Immunol. 183, 3569–3573 (2009).
Pichlmair, A. et al. RIG-I-mediated antiviral responses to single-stranded RNA bearing 5′-phosphates. Science 314, 997–1001 (2006).
Yount, J.S., Gitlin, L., Moran, T.M. & Lopez, C.B. MDA5 participates in the detection of paramyxovirus infection and is essential for the early activation of dendritic cells in response to Sendai Virus defective interfering particles. J. Immunol. 180, 4910–4918 (2008).
Yount, J.S., Kraus, T.A., Horvath, C.M., Moran, T.M. & Lopez, C.B. A novel role for viral-defective interfering particles in enhancing dendritic cell maturation. J. Immunol. 177, 4503–4513 (2006).
Acknowledgements
J.S.Y. is a postdoctoral fellow of the Irving Institute Fellowship Program of the Cancer Research Institute. Y.-Y.Y. was supported in part by the Anderson Cancer Center postdoctoral fellowship. G.C. acknowledges the Weill-Cornell/Rockefeller/Sloan-Kettering Tri-institutional Program in Chemical Biology. The authors would like to thank A. Li for technical support. T.M.M. is supported by grant AI041111 and C.B.L. is supported by grant AI083284-01, both from the US National Institute of Allergy and Infectious Diseases of the US National Institutes of Health. H.C.H. acknowledges support from The Rockefeller University, Ellison Medical Foundation, Irma T. Hirschl and Monique Weill-Caulier Trust, Lerner Trust and the US National Institute of Drug Abuse of the US National Institutes of Health (1R21DA025751-01).
Author information
Authors and Affiliations
Contributions
J.S.Y. conceived the study, designed and performed experiments, interpreted data and co-wrote the manuscript; Y.-Y.Y. and G.C. synthesized reagents for palmitoylome profiling studies; B.M., C.B.L. and T.M.M. provided reagents and expertise on influenza virus infections; H.C.H. conceived the study, designed experiments, interpreted data and co-wrote the manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Text and Figures
Supplementary Tables 1–4 & Supplementary Figures 1–8 (PDF 54120 kb)
Rights and permissions
About this article
Cite this article
Yount, J., Moltedo, B., Yang, YY. et al. Palmitoylome profiling reveals S-palmitoylation–dependent antiviral activity of IFITM3. Nat Chem Biol 6, 610–614 (2010). https://doi.org/10.1038/nchembio.405
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nchembio.405
This article is cited by
-
Lessons in self-defence: inhibition of virus entry by intrinsic immunity
Nature Reviews Immunology (2022)
-
Radio-detoxified LPS alters bone marrow-derived extracellular vesicles and endothelial progenitor cells
Stem Cell Research & Therapy (2019)
-
IFITM3 directly engages and shuttles incoming virus particles to lysosomes
Nature Chemical Biology (2019)
-
A balancing act between IFITM3 and IRF3
Cellular & Molecular Immunology (2018)
-
Antiviral Protection by IFITM3 In Vivo
Current Clinical Microbiology Reports (2018)