This article has been updated

Abstract

The poly(ADP-ribose) polymerases (PARPs) participate in many biological and pathological processes. Here we report that the PARP-13 shorter isoform (ZAPS), rather than the full-length protein (ZAP), was selectively induced by 5′-triphosphate–modified RNA (3pRNA) and functioned as a potent stimulator of interferon responses in human cells mediated by the RNA helicase RIG-I. ZAPS associated with RIG-I to promote the oligomerization and ATPase activity of RIG-I, which led to robust activation of IRF3 and NF-κB transcription factors. Disruption of the gene encoding ZAPS resulted in impaired induction of interferon-α (IFN-α), IFN-β and other cytokines after viral infection. These results indicate that ZAPS is a key regulator of RIG-I signaling during the innate antiviral immune response, which suggests its possible use as a therapeutic target for viral control.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Change history

  • 08 December 2010

    In the version of this article initially published online, the affiliation of F. Kashigi, S. Goto and S. Kameoka with the Department of Chemistry, Graduate School of Science, Hokkaido University Sapporo, Japan, was omitted. The error has been corrected for the print, PDF and HTML versions of this article.

Accessions

NCBI Reference Sequence

References

  1. 1.

    & Pattern recognition receptors and inflammation. Cell 140, 805–820 (2010).

  2. 2.

    & Cytosolic DNA recognition for triggering innate immune responses. Adv. Drug Deliv. Rev. 60, 847–857 (2008).

  3. 3.

    , , , & Innate immune modulation by RNA viruses: emerging insights from functional genomics. Nat. Rev. Immunol. 8, 644–654 (2008).

  4. 4.

    et al. The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses. Nat. Immunol. 5, 730–737 (2004).

  5. 5.

    et al. Shared and unique functions of the DExD/H-box helicases RIG-I, MDA5, and LGP2 in antiviral innate immunity. J. Immunol. 175, 2851–2858 (2005).

  6. 6.

    & RNA recognition and signal transduction by RIG-I-like receptors. Immunol. Rev. 227, 54–65 (2009).

  7. 7.

    & Innate immunity to virus infection. Immunol. Rev. 227, 75–86 (2009).

  8. 8.

    & RIGorous detection: exposing virus through RNA sensing. Science 327, 284–286 (2010).

  9. 9.

    et al. 5′-Triphosphate RNA is the ligand for RIG-I. Science 314, 994–997 (2006).

  10. 10.

    et al. RIG-I-mediated antiviral responses to single-stranded RNA bearing 5′-phosphates. Science 314, 997–1001 (2006).

  11. 11.

    et al. IPS-1, an adaptor triggering RIG-I- and Mda5-mediated type I interferon induction. Nat. Immunol. 6, 981–988 (2005).

  12. 12.

    et al. Cardif is an adaptor protein in the RIG-I antiviral pathway and is targeted by hepatitis C virus. Nature 437, 1167–1172 (2005).

  13. 13.

    , , & Identification and characterization of MAVS, a mitochondrial antiviral signaling protein that activates NF-κB and IRF 3. Cell 122, 669–682 (2005).

  14. 14.

    et al. VISA is an adapter protein required for virus-triggered IFN-β signaling. Mol. Cell 19, 727–740 (2005).

  15. 15.

    , , & RIG-I-like receptors: sensing and responding to RNA virus infection. Semin. Immunol. 21, 215–222 (2009).

  16. 16.

    et al. Reconstitution of the RIG-I pathway reveals a signaling role of unanchored polyubiquitin chains in innate immunity. Cell 141, 315–330 (2010).

  17. 17.

    et al. TRIM25 RING-finger E3 ubiquitin ligase is essential for RIG-I-mediated antiviral activity. Nature 446, 916–920 (2007).

  18. 18.

    et al. NLRC5 negatively regulates the NF-κB and type I interferon signaling pathways. Cell 141, 483–496 (2010).

  19. 19.

    , , & The expanding field of poly(ADP-ribosyl)ation reactions. 'Protein Modifications: Beyond the Usual Suspects' Review Series. EMBO Rep. 9, 1094–1100 (2008).

  20. 20.

    , & Inhibition of retroviral RNA production by ZAP, a CCCH-type zinc finger protein. Science 297, 1703–1706 (2002).

  21. 21.

    & The diverse biological roles of mammalian PARPS, a small but powerful family of poly-ADP-ribose polymerases. Front. Biosci. 13, 3046–3082 (2008).

  22. 22.

    , , & Poly(ADP-ribose): novel functions for an old molecule. Nat. Rev. Mol. Cell Biol. 7, 517–528 (2006).

  23. 23.

    et al. Regulation of Epstein-Barr virus OriP replication by poly(ADP-ribose) polymerase 1. J. Virol. 84, 4988–4997 (2010).

  24. 24.

    et al. Poly(ADP-ribose) polymerase 1 binds to Kaposi's sarcoma-associated herpesvirus (KSHV) terminal repeat sequence and modulates KSHV replication in latency. J. Virol. 78, 9936–9946 (2004).

  25. 25.

    , , , & p72 DEAD box RNA helicase is required for optimal function of the zinc-finger antiviral protein. Proc. Natl. Acad. Sci. USA 105, 4352–4357 (2008).

  26. 26.

    & ZAP-mediated mRNA degradation. RNA Biol. 5, 65–67 (2008).

  27. 27.

    , , , & Transcriptional coactivation of nuclear factor-κB-dependent gene expression by p300 is regulated by poly(ADP)-ribose polymerase-1. J. Biol. Chem. 278, 45145–45153 (2003).

  28. 28.

    et al. Resistance to endotoxic shock as a consequence of defective NF-κB activation in poly (ADP-ribose) polymerase-1 deficient mice. EMBO J. 18, 4446–4454 (1999).

  29. 29.

    , , , & B-aggressive lymphoma family proteins have unique domains that modulate transcription and exhibit poly(ADP-ribose) polymerase activity. J. Biol. Chem. 280, 33756–33765 (2005).

  30. 30.

    et al. BAL1 and BBAP are regulated by a γ interferon-responsive bidirectional promoter and are overexpressed in diffuse large B-cell lymphomas with a prominent inflammatory infiltrate. Mol. Cell. Biol. 26, 5348–5359 (2006).

  31. 31.

    et al. PARP-2 deficiency affects the survival of CD4+CD8+ double-positive thymocytes. EMBO J. 25, 4350–4360 (2006).

  32. 32.

    et al. Differential effect of PARP-2 deletion on brain injury after focal and global cerebral ischemia. J. Cereb. Blood Flow Metab. 26, 135–141 (2006).

  33. 33.

    , , , & TCDD-inducible poly(ADP-ribose) polymerase: a novel response to 2,3,7,8-tetrachlorodibenzo-p-dioxin. Biochem. Biophys. Res. Commun. 289, 499–506 (2001).

  34. 34.

    , & Immediate lymphotoxin β receptor-mediated transcriptional response in host defense against L. monocytogenes. Immunobiology 213, 353–366 (2008).

  35. 35.

    et al. Length-dependent recognition of double-stranded ribonucleic acids by retinoic acid-inducible gene-I and melanoma differentiation-associated gene 5. J. Exp. Med. 205, 1601–1610 (2008).

  36. 36.

    , & RNA polymerase III detects cytosolic DNA and induces type I interferons through the RIG-I pathway. Cell 138, 576–591 (2009).

  37. 37.

    et al. RIG-I-dependent sensing of poly(dA:dT) through the induction of an RNA polymerase III-transcribed RNA intermediate. Nat. Immunol. 10, 1065–1072 (2009).

  38. 38.

    , & Positive selection and increased antiviral activity associated with the PARP-containing isoform of human zinc-finger antiviral protein. PLoS Genet. 4, e21 (2008).

  39. 39.

    , , , & The zinc finger antiviral protein directly binds to specific viral mRNAs through the CCCH zinc finger motifs. J. Virol. 78, 12781–12787 (2004).

  40. 40.

    et al. Viral induction of the zinc finger antiviral protein is IRF3-dependent but NF-κB-independent. J. Biol. Chem. 285, 6080–6090 (2010).

  41. 41.

    , , & Antiviral defense: RIG-Ing the immune system to STING. Cytokine Growth Factor Rev. 20, 1–5 (2009).

  42. 42.

    et al. Regulation of innate antiviral defenses through a shared repressor domain in RIG-I and LGP2. Proc. Natl. Acad. Sci. USA 104, 582–587 (2007).

  43. 43.

    et al. The C-terminal regulatory domain is the RNA 5′-triphosphate sensor of RIG-I. Mol. Cell 29, 169–179 (2008).

  44. 44.

    et al. 5′-triphosphate RNA requires base-paired structures to activate antiviral signaling via RIG-I. Proc. Natl. Acad. Sci. USA 106, 12067–12072 (2009).

  45. 45.

    et al. Differential roles of MDA5 and RIG-I helicases in the recognition of RNA viruses. Nature 441, 101–105 (2006).

  46. 46.

    et al. Highly efficient endogenous human gene correction using designed zinc-finger nucleases. Nature 435, 646–651 (2005).

  47. 47.

    et al. Targeted mutagenesis in zebrafish using customized zinc-finger nucleases. Nat. Protocols 4, 1855–1867 (2009).

  48. 48.

    , & Targeted chromosomal deletions in human cells using zinc finger nucleases. Genome Res. 20, 81–89 (2010).

  49. 49.

    & Influenza B virus NS1 protein inhibits conjugation of the interferon (IFN)-induced ubiquitin-like ISG15 protein. EMBO J. 20, 362–371 (2001).

  50. 50.

    et al. Negative regulation of the RIG-I signaling by the ubiquitin ligase RNF125. Proc. Natl. Acad. Sci. USA 104, 7500–7505 (2007).

  51. 51.

    et al. The tumour suppressor CYLD is a negative regulator of RIG-I-mediated antiviral response. EMBO Rep. 9, 930–936 (2008).

  52. 52.

    , , & Riplet/RNF135, a RING finger protein, ubiquitinates RIG-I to promote interferon-beta induction during the early phase of viral infection. J. Biol. Chem. 284, 807–817 (2009).

  53. 53.

    , , , & Innate immunity induced by composition-dependent RIG-I recognition of hepatitis C virus RNA. Nature 454, 523–527 (2008).

  54. 54.

    et al. DAI (DLM-1/ZBP1) is a cytosolic DNA sensor and an activator of innate immune response. Nature 448, 501–505 (2007).

  55. 55.

    et al. Solution structures of cytosolic RNA sensor MDA5 and LGP2 C-terminal domains: identification of the RNA recognition loop in RIG-I-like receptors. J. Biol. Chem. 284, 17465–17474 (2009).

Download references

Acknowledgements

We thank T. Fujita (Kyoto University) for the luciferase reporter plasmids p-55C1BLuc and p-125Luc; J. Miyazaki (Osaka University) for the pCAGGS vector; A. Miyawaki (RIKEN) for the Venus vector; H. Kida (Hokkaido University) for NDV; A. Iwai, H. Higashi and J. Hamada for technical help; M. Yamane for the purification of human primary CD14+ monocytes; and S. Tamura and T. Moriyama for advice on recombinant protein purification. Supported by the Ministry of Education, Culture, Sports, Science and Technology of Japan (Grant-in-Aid for Young Scientists (A) to S.H., Young Scientists (S) to A.T.) and Scientific Research in Priority Areas (A.T.), IRYO HOJIN SHADAN JIKOKAI (H. Tanaka & N. Takayanagi) (A.T.), the Astellas Foundation for Research on Metabolic Disorders (A.T.), the Kanae Foundation for the Promotion of Medical Science (A.T.), the Kato Memorial Bioscience Foundation (A.T.) and the Yasuda Medical Foundation (A.T.).

Author information

Author notes

    • Sumio Hayakawa
    • , Souichi Shiratori
    •  & Hiroaki Yamato

    These authors contributed equally to this work.

Affiliations

  1. Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Hokkaido University, Sapporo, Japan.

    • Sumio Hayakawa
    • , Souichi Shiratori
    • , Hiroaki Yamato
    • , Takeshi Kameyama
    • , Chihiro Kitatsuji
    • , Fumi Kashigi
    • , Showhey Goto
    • , Shoichiro Kameoka
    • , Taisho Yamada
    • , Mika Kazumata
    • , Maiko Sato
    •  & Akinori Takaoka
  2. Department of Hematology and Oncology, Hokkaido University Graduate School of Medicine, Sapporo, Japan.

    • Souichi Shiratori
    • , Junji Tanaka
    •  & Masahiro Imamura
  3. Department of Gastroenterology, Hokkaido University Graduate School of Medicine, Sapporo, Japan.

    • Hiroaki Yamato
    •  & Masahiro Asaka
  4. Department of Bioresources, Hokkaido University Research Center for Zoonosis Control, Sapporo, Japan.

    • Daisuke Fujikura
    •  & Tadaaki Miyazaki
  5. Laboratory of Pathophysiology and Signal Transduction, Hokkaido University Graduate School of Medicine, Sapporo, Japan.

    • Tatsuaki Mizutani
    •  & Yusuke Ohba
  6. Research Center of Infection-Associated Cancer, Institute for Genetic Medicine, Hokkaido University, Sapporo, Japan.

    • Akinori Takaoka
  7. Department of Chemistry, Graduate School of Science, Hokkaido University, Sapporo, Japan.

    • Fumi Kashigi
    • , Showhey Goto
    •  & Shoichiro Kameoka

Authors

  1. Search for Sumio Hayakawa in:

  2. Search for Souichi Shiratori in:

  3. Search for Hiroaki Yamato in:

  4. Search for Takeshi Kameyama in:

  5. Search for Chihiro Kitatsuji in:

  6. Search for Fumi Kashigi in:

  7. Search for Showhey Goto in:

  8. Search for Shoichiro Kameoka in:

  9. Search for Daisuke Fujikura in:

  10. Search for Taisho Yamada in:

  11. Search for Tatsuaki Mizutani in:

  12. Search for Mika Kazumata in:

  13. Search for Maiko Sato in:

  14. Search for Junji Tanaka in:

  15. Search for Masahiro Asaka in:

  16. Search for Yusuke Ohba in:

  17. Search for Tadaaki Miyazaki in:

  18. Search for Masahiro Imamura in:

  19. Search for Akinori Takaoka in:

Contributions

S.H., S.S., H.Y., T.K., C.K., F.K., S.G., S.K., T.Y., M.K., M.S., J.T., M.A. and M.I. planned studies, did experiments and analyzed data; D.F. and T. Miyazaki contributed to viral infection experiments and helped with data analyses; T. Mizutani and Y.O. did fluorescence microscopy experiments and FRET analysis; and A.T. supervised the project, designed experiments and wrote the manuscript with comments from the coauthors.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Akinori Takaoka.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–5 and Tables 1–2

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ni.1963

Further reading