Article

Secreted tryptophanyl-tRNA synthetase as a primary defence system against infection

  • Nature Microbiology 2, Article number: 16191 (2016)
  • doi:10.1038/nmicrobiol.2016.191
  • Download Citation
Received:
Accepted:
Published online:

Abstract

The N-terminal truncated form of a protein synthesis enzyme, tryptophanyl-tRNA synthetase (mini-WRS), is secreted as an angiostatic ligand. However, the secretion and function of the full-length WRS (FL-WRS) remain unknown. Here, we report that the FL-WRS, but not mini-WRS, is rapidly secreted upon pathogen infection to prime innate immunity. Blood levels of FL-WRS were increased in sepsis patients, but not in those with sterile inflammation. FL-WRS was secreted from monocytes and directly bound to macrophages via a toll-like receptor 4 (TLR4)–myeloid differentiation factor 2 (MD2) complex to induce phagocytosis and chemokine production. Administration of FL-WRS into Salmonella typhimurium-infected mice reduced the levels of bacteria and improved mouse survival, whereas its titration with the specific antibody aggravated the infection. The N-terminal 154-amino-acid eukaryote-specific peptide of WRS was sufficient to recapitulate FL-WRS activity and its interaction mode with TLR4–MD2 is now suggested. Based on these results, secretion of FL-WRS appears to work as a primary defence system against infection, acting before full activation of innate immunity.

  • Subscribe to Nature Microbiology for full access:

    $59

    Subscribe

Additional access options:

Already a subscriber?  Log in  now or  Register  for online access.

References

  1. 1.

    et al. HMG-1 as a late mediator of endotoxin lethality in mice. Science 285, 248–251 (1999).

  2. 2.

    et al. Reversing established sepsis with antagonists of endogenous high-mobility group box 1. Proc. Natl Acad. Sci. USA 101, 296–301 (2004).

  3. 3.

    et al. HSP70 as endogenous stimulus of the Toll/interleukin-1 receptor signal pathway. J. Biol. Chem. 277, 15107–15112 (2002).

  4. 4.

    , & Extracellular activities of aminoacyl-tRNA synthetases: new mediators for cell–cell communication. Top. Curr. Chem. 344, 145–166 (2014).

  5. 5.

    , & New functions of aminoacyl-tRNA synthetases beyond translation. Nat. Rev. Mol. Cell Biol. 11, 668–674 (2010).

  6. 6.

    , , & Transcriptional regulation of the interferon-gamma-inducible tryptophanyl-tRNA synthetase includes alternative splicing. J. Biol. Chem. 270, 397–403 (1995).

  7. 7.

    et al. A human aminoacyl-tRNA synthetase as a regulator of angiogenesis. Proc. Natl Acad. Sci. USA 99, 173–177 (2002).

  8. 8.

    et al. Comparative proteomic analysis reveals activation of mucosal innate immune signaling pathways during cholera. Infect. Immun. 83, 1089–1103 (2015).

  9. 9.

    , , , & Cellular gene expression altered by human cytomegalovirus: global monitoring with oligonucleotide arrays. Proc. Natl Acad. Sci. USA 95, 14470–14475 (1998).

  10. 10.

    , , & Genomic analysis of the host response to hepatitis B virus infection. Proc. Natl Acad. Sci. USA 101, 6669–6674 (2004).

  11. 11.

    , , & In vivo high spatiotemporal resolution visualization of circulating T lymphocytes in high endothelial venules of lymph nodes. J. Biomed. Opt. 18, 036005 (2013).

  12. 12.

    , , , & Insertion of enhanced green fluorescent protein into the lysozyme gene creates mice with green fluorescent granulocytes and macrophages. Blood 96, 719–726 (2000).

  13. 13.

    , , & In vivo quantitation of injected circulating tumor cells from great saphenous vein based on video-rate confocal microscopy. Biomed. Opt. Express 6, 2158–2167 (2015).

  14. 14.

    , , & Construction of a large synthetic human scFv library with six diversified CDRs and high functional diversity. Mol. Cells 27, 225–235 (2009).

  15. 15.

    et al. Differential roles of TLR2 and TLR4 in recognition of Gram-negative and Gram-positive bacterial cell wall components. Immunity 11, 443–451 (1999).

  16. 16.

    et al. Mal (MyD88-adapter-like) is required for Toll-like receptor-4 signal transduction. Nature 413, 78–83 (2001).

  17. 17.

    et al. The structural basis of lipopolysaccharide recognition by the TLR4–MD-2 complex. Nature 458, 1191–1195 (2009).

  18. 18.

    et al. MD-2 is required for disulfide HMGB1-dependent TLR4 signaling. J. Exp. Med. 212, 5–14 (2015).

  19. 19.

    , & An unusual dimeric structure and assembly for TLR4 regulator RP105-MD-1. Nat. Struct. Mol. Biol. 18, 1028–1035 (2011).

  20. 20.

    et al. Crystal structures that suggest late development of genetic code components for differentiating aromatic side chains. Proc. Natl Acad. Sci. USA 100, 15376–15380 (2003).

  21. 21.

    , & HMGB1 a multifunctional alarmin driving autoimmune and inflammatory disease. Nat. Rev. Rheumatol. 8, 195–202 (2012).

  22. 22.

    et al. Core temperature correlates with expression of selected stress and immunomodulatory genes in febrile patients with sepsis and noninfectious SIRS. Cell Stress Chaperones 15, 55–66 (2010).

  23. 23.

    et al. Synergy and cross-tolerance between toll-like receptor (TLR) 2- and TLR4-mediated signaling pathways. J. Immunol. 165, 7096–7101 (2000).

  24. 24.

    et al. Orthogonal use of a human tRNA synthetase active site to achieve multifunctionality. Nat. Struct. Mol. Biol. 17, 57–61 (2010).

  25. 25.

    et al. Definitions for sepsis and organ failure and guidelines for the use of innovative therapies in sepsis. The ACCP/SCCM Consensus Conference Committee. American College of Chest Physicians/Society of Critical Care Medicine. Chest 101, 1644–1655 (1992).

  26. 26.

    et al. Classification criteria for Sjogren's syndrome: a revised version of the European criteria proposed by the American-European Consensus Group. Ann. Rheum. Dis. 61, 554–558 (2002).

  27. 27.

    et al. American College of Rheumatology classification criteria for Sjogren's syndrome: a data-driven, expert consensus approach in the Sjogren's International Collaborative Clinical Alliance cohort. Arthritis Care Res. 64, 475–487 (2012).

  28. 28.

    et al. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum. 31, 315–324 (1988).

  29. 29.

    et al. 2010 Rheumatoid Arthritis Classification Criteria: an American College of Rheumatology/European League Against Rheumatism collaborative initiative. Ann. Rheum. Dis. 69, 1580–1588 (2010).

  30. 30.

    et al. Human lysyl-tRNA synthetase is secreted to trigger proinflammatory response. Proc. Natl Acad. Sci. USA 102, 6356–6361 (2005).

  31. 31.

    et al. Pheophytin a and chlorophyll a suppress neuroinflammatory responses in lipopolysaccharide and interferon-γ-stimulated BV2 microglia. Life Sci. 103, 59–67 (2014).

  32. 32.

    et al. Essential role of MD-2 in LPS responsiveness and TLR4 distribution. Nat. Immunol. 3, 667–672 (2002).

  33. 33.

    et al. Cutting edge: Toll-like receptor 4 (TLR4)-deficient mice are hyporesponsive to lipopolysaccharide: evidence for TLR4 as the Lps gene product. J. Immunol. 162, 3749–3752 (1999).

  34. 34.

    , & Cutting edge: TLR2-deficient and MyD88-deficient mice are highly susceptible to Staphylococcus aureus infection. J. Immunol. 165, 5392–5396 (2000).

Download references

Acknowledgements

This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2010-0012505), Bio-Synergy/research Project (2014M3A9C4066465) and Global Frontier Project grants nos. NRF-M3A6A4-2010-0029785 and 2015M3A6A4065732 of the National Research Foundation funded by the Ministry of Science, ICT & Future Planning (MSIP) of Korea.

Author information

Author notes

    • Young Ha Ahn
    •  & Sunyoung Park

    These authors contributed equally to this work.

Affiliations

  1. College of Pharmacy, Seoul National University, Seoul 08826, Republic of Korea

    • Young Ha Ahn
    • , Joo-Youn Lee
    •  & Sunghoon Kim
  2. College of Korean Medicine, Daejeon University, Daejeon 34520, Republic of Korea

    • Sunyoung Park
    • , Jeong June Choi
    • , Bo-Kyung Park
    •  & Mirim Jin
  3. Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul 08826, Republic of Korea

    • Kyung Hee Rhee
    •  & Byung Woo Han
  4. Medicinal Bioconvergence Research Center, Seoul National University, Suwon 16229, Republic of Korea

    • Eunjoo Kang
    • , Nam Hoon Kwon
    •  & Sunghoon Kim
  5. Graduate School of Nanoscience and Technology, Korea Advanced Institute of Science and Technology, Daejeon 34141, Republic of Korea

    • Soyeon Ahn
    •  & Pilhan Kim
  6. Laboratory Animal Resource Center, Korea Research Institute of Bioscience and Biotechnology, University of Science and Technology, Daejeon 34113, Republic of Korea

    • Chul-Ho Lee
  7. College of Veterinary Medicine, Chungnam National University, Daejeon 34134, Republic of Korea

    • Jong Soo Lee
  8. Department of Pharmaceutical Science, College of Pharmacy, Kyung Hee University, Seoul 02447, Republic of Korea

    • Kyung-Soo Inn
  9. Divison of Rheumatology, Department of Internal Medicine, Seoul St Mary's Hospital, The Catholic University of Korea, Seoul 06591, Republic of Korea

    • Mi-La Cho
  10. The Rheumatism Research Center, Catholic Research Institute of Medical Science, The Catholic University of Korea, Seoul 06591, Republic of Korea

    • Sung-Hwan Park
  11. Division of Allergy and Immunology, Department of Internal Medicine, Yonsei University College of Medicine, Seoul 03722, Republic of Korea

    • Kyunghee Park
    • , Hye Jung Park
    • , Jae-Hyun Lee
    •  & Jung-Won Park
  12. Institute of Allergy, Yonsei University College of Medicine, Seoul 03722, Republic of Korea

    • Kyunghee Park
    • , Hye Jung Park
    • , Jae-Hyun Lee
    •  & Jung-Won Park
  13. Departments of Bioinspired Science and Life Science, Ewha Womans University, Seoul 03760, Republic of Korea

    • Hyunbo Shim
  14. Korea Chemical Bank, Korea Research Institute of Chemical Technology, Daejeon 34114, Republic of Korea

    • Joo-Youn Lee
  15. College of Pharmacy, Korea University, Sejong 30019, Republic of Korea

    • Youngho Jeon
  16. Department of Pulmonary and Critical Care Medicine, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of Korea

    • Jin Won Huh

Authors

  1. Search for Young Ha Ahn in:

  2. Search for Sunyoung Park in:

  3. Search for Jeong June Choi in:

  4. Search for Bo-Kyung Park in:

  5. Search for Kyung Hee Rhee in:

  6. Search for Eunjoo Kang in:

  7. Search for Soyeon Ahn in:

  8. Search for Chul-Ho Lee in:

  9. Search for Jong Soo Lee in:

  10. Search for Kyung-Soo Inn in:

  11. Search for Mi-La Cho in:

  12. Search for Sung-Hwan Park in:

  13. Search for Kyunghee Park in:

  14. Search for Hye Jung Park in:

  15. Search for Jae-Hyun Lee in:

  16. Search for Jung-Won Park in:

  17. Search for Nam Hoon Kwon in:

  18. Search for Hyunbo Shim in:

  19. Search for Byung Woo Han in:

  20. Search for Pilhan Kim in:

  21. Search for Joo-Youn Lee in:

  22. Search for Youngho Jeon in:

  23. Search for Jin Won Huh in:

  24. Search for Mirim Jin in:

  25. Search for Sunghoon Kim in:

Contributions

Co-first authors Y.H.A. and S.P. performed the experiments and analysed the data. J.J.C., B.-K.P., K.H.R., E.K., S.A., K.-S.I., N.H.K., H.S., B.W.H., P.K., J.-Y.L. and Y.J. performed experiments. C.-H.L. and J.S.L. provided knockout animals. M.-L.C., S.-H.P., K.P., H.J.P., J.-H.L., J.-W.P. and J.W.H. provided sera and discussed clinical data. M.J. and S.K. designed the experiments, supervised the research and wrote the paper. All authors approved the final manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Mirim Jin or Sunghoon Kim.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary Figures 1–9, Supplementary Tables 1 and 2, legends for Supplementary Videos 1–6

Videos

  1. 1.

    Supplementary Video 1

    Infiltration of neutrophil and monocyte/macraophage 30 min after PBS injection.

  2. 2.

    Supplementary Video 2

    Infiltration of neutrophil and monocyte/macraophage 4 hr after PBS injection.

  3. 3.

    Supplementary Video 3

    Infiltration of neutrophil and monocyte/macraophage 30 min after FL-WRS injection.

  4. 4.

    Supplementary Video 4

    Infiltration of neutrophil and monocyte/macraophage 4 hr after FL-WRS injection.

  5. 5.

    Supplementary Video 5

    Infiltration of neutrophil and monocyte/macraophage 30 min after mini-WRS 23 injection.

  6. 6.

    Supplementary Video 6

    Infiltration of neutrophil and monocyte/macraophage 4 hr after mini-WRS injection.