The DEAH-Box RNA Helicase DHX15 Activates NF-κB and MAPK Signaling Downstream of MAVS During Antiviral Responses
Selecting the Antiviral Response with an RNA Helicase
The RIG-I–like receptor family of RNA helicases detect viral RNA, activate the mitochondrial adaptor protein MAVS, and stimulate mitogen-activated protein kinase (MAPK) and nuclear factor κB (NF-κB) signaling, as well as interferon production. Mosallanejad et al. found that another RNA helicase, DHX15, physically associated with MAVS through its RNA helicase domain to facilitate the activation of MAPK and NF-κB signaling, but not interferon production, in cells treated with an RNA mimetic or infected with RNA viruses. Drosophila DHX15 also activated MAPKs, suggesting the DHX15 may play a more general role in stress responses.
Abstract
During infection with an RNA virus, the DExD/H-box RNA helicases RIG-I (retinoic acid–inducible gene I) and MDA5 (melanoma differentiation–associated gene 5) activate the interferon regulatory factor 3 (IRF3), nuclear factor κB (NF-κB), c-Jun amino-terminal kinase (JNK), and p38 mitogen-activated protein kinase (MAPK) signaling pathways through an unknown mechanism involving the adaptor protein MAVS (mitochondrial antiviral signaling). We used a Drosophila misexpression screen to identify DEAH-box polypeptide 15 (DHX15) as an activator of the p38 MAPK pathway. Human DHX15 contributed to the activation of the NF-κB, JNK, and p38 MAPK pathways, but not the IRF3 pathway, in response to the synthetic double-stranded RNA analog poly(I:C) (polyinosinic-polycytidylic acid), and DHX15 was required for optimal cytokine production in response to poly(I:C) and infection with RNA virus. DHX15 physically interacted with MAVS and mediated the MAVS-dependent activation of the NF-κB and MAPK pathways. Furthermore, DHX15 was required for poly(I:C)- and RNA virus–dependent, MAVS-mediated apoptosis. Thus, our findings indicate that, in RIG-I–like receptor signaling, DHX15 specifically stimulates the NF-κB and MAPK pathways downstream of MAVS and contributes to MAVS-mediated cytokine production and apoptosis.
Get full access to this article
View all available purchase options and get full access to this article.
Already a Subscriber?Sign In
Supplementary Material
Summary
Fig. S1. A misexpression screen for activators of the p38 MAPK pathway in Drosophila.
Fig. S2. Amino acid alignment of human, fly, and yeast DHX15 proteins.
Fig. S3. DHX15 is required for optimal poly(I:C)-induced cytokine production.
Fig. S4. The ATPase activity of DHX15 is not required for DHX15-mediated activation of the MAPK pathway.
Fig. S5. The relationship between DHX15, MAVS, and TRAF6 in RLR signaling.
Fig. S6. DHX15 is required for poly(I:C)-induced apoptosis.
Fig. S7. DHX15 is required for the SeV-induced production of IFN-β by HEK 293 cells.
Fig. S8. Schematic model of the proposed roles of DHX15 in the antiviral response.
Resources
File (7_ra40_sm.pdf)
REFERENCES AND NOTES
1
Janeway C. A., Medzhitov R., Innate immune recognition. Annu. Rev. Immunol. 20, 197–216 (2002).
2
Jensen S., Thomsen A. R., Sensing of RNA viruses: A review of innate immune receptors involved in recognizing RNA virus invasion. J. Virol. 86, 2900–2910 (2012).
3
Schmidt A., Endres S., Rothenfusser S., Pattern recognition of viral nucleic acids by RIG-I-like helicases. J. Mol. Med. 89, 5–12 (2011).
4
Takeuchi O., Akira S., Pattern recognition receptors and inflammation. Cell 140, 805–820 (2010).
5
Yoneyama M., Kikuchi M., Natsukawa T., Shinobu N., Imaizumi T., Miyagishi M., Taira K., Akira S., Fujita T., The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses. Nat. Immunol. 5, 730–737 (2004).
6
Yoneyama M., Kikuchi M., Matsumoto K., Imaizumi T., Miyagishi M., Taira K., Foy E., Loo Y. M., Gale M., Akira S., Yonehara S., Kato A., Fujita T., 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).
7
Kato H., Takeuchi O., Mikamo-Satoh E., Hirai R., Kawai T., Matsushita K., Hiiragi A., Dermody T. S., Fujita T., Akira S., 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).
8
Seth R. B., Sun L., Ea C. K., Chen Z. J., Identification and characterization of MAVS, a mitochondrial antiviral signaling protein that activates NF-κB and IRF3. Cell 122, 669–682 (2005).
9
Kawai T., Takahashi K., Sato S., Coban C., Kumar H., Kato H., Ishii K. J., Takeuchi O., Akira S., IPS-1, an adaptor triggering RIG-I- and Mda5-mediated type I interferon induction. Nat. Immunol. 6, 981–988 (2005).
10
Xu L. G., Wang Y. Y., Han K. J., Li L. Y., Zhai Z., Shu H. B., VISA is an adapter protein required for virus-triggered IFN-β signaling. Mol. Cell 19, 727–740 (2005).
11
Meylan E., Curran J., Hofmann K., Moradpour D., Binder M., Bartenschlager R., Tschopp J., Cardif is an adaptor protein in the RIG-I antiviral pathway and is targeted by hepatitis C virus. Nature 437, 1167–1172 (2005).
12
Saha S. K., Pietras E. M., He J. Q., Kang J. R., Liu S. Y., Oganesyan G., Shahangian A., Zarnegar B., Shiba T. L., Wang Y., Cheng G., Regulation of antiviral responses by a direct and specific interaction between TRAF3 and Cardif. EMBO J. 25, 3257–3263 (2006).
13
Tang E. D., Wang C. Y., TRAF5 is a downstream target of MAVS in antiviral innate immune signaling. PLOS One 5, e9172 (2010).
14
Yoshida R., Takaesu G., Yoshida H., Okamoto F., Yoshioka T., Choi Y., Akira S., Kawai T., Yoshimura A., Kobayashi T., TRAF6 and MEKK1 play a pivotal role in the RIG-I-like helicase antiviral pathway. J. Biol. Chem. 283, 36211–36220 (2008).
15
Oshiumi H., Sakai K., Matsumoto M., Seya T., DEAD/H BOX 3 (DDX3) helicase binds the RIG-I adaptor IPS-1 to up-regulate IFN-β-inducing potential. Eur. J. Immunol. 40, 940–948 (2010).
16
Schröder M., Baran M., Bowie A. G., Viral targeting of DEAD box protein 3 reveals its role in TBK1/IKKε-mediated IRF activation. EMBO J. 27, 2147–2157 (2008).
17
Soulat D., Bürckstümmer T., Westermayer S., Goncalves A., Bauch A., Stefanovic A., Hantschel O., Bennett K. L., Decker T., Superti-Furga G., The DEAD-box helicase DDX3X is a critical component of the TANK-binding kinase 1-dependent innate immune response. EMBO J. 27, 2135–2146 (2008).
18
Miyashita M., Oshiumi H., Matsumoto M., Seya T., DDX60, a DEXD/H box helicase, is a novel antiviral factor promoting RIG-I-like receptor-mediated signaling. Mol. Cell. Biol. 31, 3802–3819 (2011).
19
Mitoma H., Hanabuchi S., Kim T., Bao M., Zhang Z., Sugimoto N., Liu Y. J., The DHX33 RNA helicase senses cytosolic RNA and activates the NLRP3 inflammasome. Immunity 39, 123–135 (2013).
20
Zhang Z., Kim T., Bao M., Facchinetti V., Jung S. Y., Ghaffari A. A., Qin J., Cheng G., Liu Y. J., DDX1, DDX21, and DHX36 helicases form a complex with the adaptor molecule TRIF to sense dsRNA in dendritic cells. Immunity 34, 866–878 (2011).
21
Zhang Z., Yuan B., Lu N., Facchinetti V., Liu Y. J., DHX9 pairs with IPS-1 to sense double-stranded RNA in myeloid dendritic cells. J. Immunol. 187, 4501–4508 (2011).
22
Besch R., Poeck H., Hohenauer T., Senft D., Häcker G., Berking C., Hornung V., Endres S., Ruzicka T., Rothenfusser S., Hartmann G., Proapoptotic signaling induced by RIG-I and MDA-5 results in type I interferon–independent apoptosis in human melanoma cells. J. Clin. Invest. 119, 2399–2411 (2009).
23
Chattopadhyay S., Marques J. T., Yamashita M., Peters K. L., Smith K., Desai A., Williams B. R. G., Sen G. C., Viral apoptosis is induced by IRF-3-mediated activation of Bax. EMBO J. 29, 1762–1773 (2010).
24
Matsuzawa A., Saegusa K., Noguchi T., Sadamitsu C., Nishitoh H., Nagai S., Koyasu S., Matsumoto K., Takeda K., Ichijo H., ROS-dependent activation of the TRAF6-ASK1-p38 pathway is selectively required for TLR4-mediated innate immunity. Nat. Immunol. 6, 587–592 (2005).
25
Sekine Y., Hatanaka R., Watanabe T., Sono N., Iemura S., Natsume T., Kuranaga E., Miura M., Takeda K., Ichijo H., The kelch repeat protein KLHDC10 regulates oxidative stress-induced ASK1 activation by suppressing PP5. Mol. Cell 48, 692–704 (2012).
26
Sekine Y., Takagahara S., Hatanaka R., Watanabe T., Oguchi H., Noguchi T., Naguro I., Kobayashi K., Tsunoda M., Funatsu T., Nomura H., Toyoda T., Matsuki N., Kuranaga E., Miura M., Takeda K., Ichijo H., p38 MAPKs regulate the expression of genes in the dopamine synthesis pathway through phosphorylation of NR4A nuclear receptors. J. Cell Sci. 124, 3006–3016 (2011).
27
Zhuang Z. H., Zhou Y., Yu M. C., Silverman N., Ge B. X., Regulation of Drosophila p38 activation by specific MAP2 kinase and MAP3 kinase in response to different stimuli. Cell. Signal. 18, 441–448 (2006).
28
Rocak S., Linder P., DEAD-box proteins: The driving forces behind RNA metabolism. Nat. Rev. Mol. Cell Biol. 5, 232–241 (2004).
29
Fouraux M. A., Kolkman M. J. M., der Heijden A. V., Jong A. S. D., Venrooij W. J. V., Pruijn G. J. M., The human La (SS-B) autoantigen interacts with DDX15/hPrp43, a putative DEAH-box RNA helicase. RNA 8, 1428–1443 (2002).
30
Niu Z., Jin W., Zhang L., Li X., Tumor suppressor RBM5 directly interacts with the DExD/H-box protein DHX15 and stimulates its helicase activity. FEBS Lett. 586, 977–983 (2012).
31
Tariq A., Garncarz W., Handl C., Balik A., Pusch O., Jantsch M. F., RNA-interacting proteins act as site-specific repressors of ADAR2-mediated RNA editing and fluctuate upon neuronal stimulation. Nucleic Acids Res. 41, 2581–2593 (2013).
32
Panne D., Maniatis T., Harrison S. C., An atomic model of the interferon-β enhanceosome. Cell 129, 1111–1123 (2007).
33
Panne D., The enhanceosome. Curr. Opin. Struct. Biol. 18, 236–242 (2008).
34
Arenas J. E., Abelson J. N., Prp43: An RNA helicase-like factor involved in spliceosome disassembly. Proc. Natl. Acad. Sci. U.S.A. 94, 11798–11802 (1997).
35
Combs D. J., Nagel R. J., Ares M., Stevens S. W., Prp43p is a DEAH-box spliceosome disassembly factor essential for ribosome biogenesis. Mol. Cell. Biol. 26, 523–534 (2006).
36
Yoneyama M., Suhara W., Fukuhara Y., Sato M., Ozato K., Fujita T., Autocrine amplification of type I interferon gene expression mediated by interferon stimulated gene factor 3 (ISGF3). J. Biochem. 120, 160–169 (1996).
37
Scott I., Degradation of RIG-I following cytomegalovirus infection is independent of apoptosis. Microbes Infect. 11, 973–979 (2009).
38
Lei Y., Moore C. B., Liesman R. M., O’Connor B. P., Bergstralh D. T., Chen Z. J., Pickles R. J., Ting J. P. Y., MAVS-mediated apoptosis and its inhibition by viral proteins. PLOS One 4, e5466 (2009).
39
Mikkelsen S. S., Jensen S. B., Chiliveru S., Melchjorsen J., Julkunen I., Gaestel M., Arthur J. S. C., Flavell R. A., Ghosh S., Paludan S. R., RIG-I-mediated activation of p38 MAPK is essential for viral induction of interferon and activation of dendritic cells: Dependence on TRAF2 and TAK1. J. Biol. Chem. 284, 10774–10782 (2009).
40
Wang J., Basagoudanavar S. H., Wang X., Hopewell E., Albrecht R., García-Sastre A., Balachandran S., Beg A. A., NF-κB RelA subunit is crucial for early IFN-β expression and resistance to RNA virus replication. J. Immunol. 185, 1720–1729 (2010).
41
Calleja M., Herranz H., Estella C., Casal J., Lawrence P., Simpson P., Morata G., Generation of medial and lateral dorsal body domains by the pannier gene of Drosophila. Development 127, 3971–3980 (2000).
42
Adachi-Yamada T., Nakamura M., Irie K., Tomoyasu Y., Sano Y., Mori E., Goto S., Ueno N., Nishida Y., Matsumoto K., p38 mitogen-activated protein kinase can be involved in transforming growth factor β superfamily signal transduction in Drosophila wing morphogenesis. Mol. Cell. Biol. 19, 2322–2329 (1999).
43
Ninomiya-Tsuji J., Kishimoto K., Hiyama A., Inoue J., Cao Z., Matsumoto K., The kinase TAK1 can activate the NIK-IκB as well as the MAP kinase cascade in the IL-1 signalling pathway. Nature 398, 252–256 (1999).
44
Kumagai K., Kojima H., Okabe T., Nagano T., Development of a highly sensitive, high-throughput assay for glycosyltransferases using enzyme-coupled fluorescence detection. Anal. Biochem. 447, 146–155 (2014).
45
Onoguchi K., Onomoto K., Takamatsu S., Jogi M., Takemura A., Morimoto S., Julkunen I., Namiki H., Yoneyama M., Fujita T., Virus-infection or 5′ppp-RNA activates antiviral signal through redistribution of IPS-1 mediated by MFN1. PLOS Pathog. 6, e1001012 (2010).
46
Ishida T., Mizushima S., Azuma S., Kobayashi N., Tojo T., Suzuki K., Aizawa S., Watanabe T., Mosialos G., Kieff E., Yamamoto T., Inoue J., Identification of TRAF6, a novel tumor necrosis factor receptor-associated factor protein that mediates signaling from an amino-terminal domain of the CD40 cytoplasmic region. J. Biol. Chem. 271, 28745–28748 (1996).
Information & Authors
Information
Published In
Science Signaling
Volume 7 | Issue 323
April 2014
April 2014
Copyright
Copyright © 2014, American Association for the Advancement of Science.
Submission history
Received: 22 October 2013
Accepted: 11 April 2014
Acknowledgments
We thank T. Adachi-Yamada, S. B. Carroll, Y. Hiromi, the Bloomington Drosophila Stock Center, the Drosophila Genetic Resource Center at Kyoto Institute of Technology, and the NIG-FLY Stock Center for fly strains and materials; T. Fujita for EMCV and plasmids; and H. Nakano, J.-i. Inoue, and K. Matsumoto for plasmids. We thank T. Okazaki, Y. Gotoh, E. Kuranaga, and M. Miura for giving crucial advice, support, and materials. We are grateful to all the members of the Laboratory of Cell Signaling for their crucial comments. Funding: This work was supported by KAKENHI from the Japan Society for the Promotion of Science and the Ministry of Education, Culture, Sports, Science and Technology (MEXT); Global Center of Excellence Program; the “Understanding of molecular and environmental bases for brain health” conducted under the Strategic Research Program for Brain Sciences by MEXT; the Naito Foundation Natural Science Scholarship; the Cosmetology Research Foundation; and the Tokyo Biochemical Research Foundation. Author contributions: K.M., Y.S., K.T., and H.I. designed the study; K.M., Y.S., S.I.-K., A.M., and I.N. performed the experiments and analyzed the data; K.M., Y.S., and H.I. wrote the manuscript; and K.K. and T.N. gave technical support and conceptual advice for measurement of the ATPase activity of DHX15. Competing interests: The authors declare that they have no competing interests.
Authors
Metrics & Citations
Metrics
Article Usage
Altmetrics
Citations
Export citation
Select the format you want to export the citation of this publication.
Cited by
- DHX15 is required to control RNA virus-induced intestinal inflammation, Cell Reports, 35, 12, (109205), (2021).https://doi.org/10.1016/j.celrep.2021.109205
- The RNA helicase Dhx15 mediates Wnt-induced antimicrobial protein expression in Paneth cells, Proceedings of the National Academy of Sciences, 118, 4, (e2017432118), (2021).https://doi.org/10.1073/pnas.2017432118
- Identifying Infliximab- (IFX-) Responsive Blood Signatures for the Treatment of Rheumatoid Arthritis, BioMed Research International, 2021, (1-10), (2021).https://doi.org/10.1155/2021/5556784
- Regulation of RNA helicase activity: principles and examples, Biological Chemistry, 402, 5, (529-559), (2021).https://doi.org/10.1515/hsz-2020-0362
- The loss of DHX15 impairs endothelial energy metabolism, lymphatic drainage and tumor metastasis in mice, Communications Biology, 4, 1, (2021).https://doi.org/10.1038/s42003-021-02722-w
- Nlrp6 regulates intestinal antiviral innate immunity, Science, 350, 6262, (826-830), (2021)./doi/10.1126/science.aab3145
- Deciphering the Fine-Tuning of the Retinoic Acid-Inducible Gene-I Pathway in Teleost Fish and Beyond, Frontiers in Immunology, 12, (2021).https://doi.org/10.3389/fimmu.2021.679242
- DHX15 Inhibits Autophagy and the Proliferation of Hepatoma Cells, Frontiers in Medicine, 7, (2021).https://doi.org/10.3389/fmed.2020.591736
- Viral infection-induced changes in the expression profile of non-RLR DExD/H-box RNA helicases (DDX1, DDX3, DHX9, DDX21 and DHX36) in zebrafish and common carp, Fish & Shellfish Immunology, 104, (62-73), (2020).https://doi.org/10.1016/j.fsi.2020.06.010
- Distinct and Orchestrated Functions of RNA Sensors in Innate Immunity, Immunity, 53, 1, (26-42), (2020).https://doi.org/10.1016/j.immuni.2020.03.017
- See more
Loading...
View Options
Get Access
Log in to view the full text
AAAS login provides access to Science for AAAS Members, and access to other journals in the Science family to users who have purchased individual subscriptions.
- Become a AAAS Member
- Activate your AAAS ID
- Purchase Access to Other Journals in the Science Family
- Account Help
Log in via OpenAthens.
Log in via Shibboleth.
More options
Register for free to read this article
As a service to the community, this article is available for free. Login or register for free to read this article.
View options
PDF format
Download this article as a PDF file
Download PDF