DHX36 Enhances RIG-I Signaling by Facilitating PKR-mediated Antiviral Stress Granule Formation

Overview

Journal PLoS Pathog

Specialty Microbiology

Date 2014 Mar 22

PMID 24651521

Citations 94

Authors

Ji-Seung Yoo

Kiyohiro Takahasi

Chen Seng Ng

Ryota Ouda

Koji Onomoto

Mitsutoshi Yoneyama

Janice Ching Lai

Simon Lattmann

Yoshikuni Nagamine

Tadashi Matsui

Kuniyoshi Iwabuchi

Hiroki Kato

Takashi Fujita

Affiliations

Soon will be listed here.

Abstract

RIG-I is a DExD/H-box RNA helicase and functions as a critical cytoplasmic sensor for RNA viruses to initiate antiviral interferon (IFN) responses. Here we demonstrate that another DExD/H-box RNA helicase DHX36 is a key molecule for RIG-I signaling by regulating double-stranded RNA (dsRNA)-dependent protein kinase (PKR) activation, which has been shown to be essential for the formation of antiviral stress granule (avSG). We found that DHX36 and PKR form a complex in a dsRNA-dependent manner. By forming this complex, DHX36 facilitates dsRNA binding and phosphorylation of PKR through its ATPase/helicase activity. Using DHX36 KO-inducible MEF cells, we demonstrated that DHX36 deficient cells showed defect in IFN production and higher susceptibility in RNA virus infection, indicating the physiological importance of this complex in host defense. In summary, we identify a novel function of DHX36 as a critical regulator of PKR-dependent avSG to facilitate viral RNA recognition by RIG-I-like receptor (RLR).

Citing Articles

Established and Emerging Roles of DEAD/H-Box Helicases in Regulating Infection and Immunity.

Parthun M, Long M, Hemann E Immunol Rev. 2024; 329(1):e13426.

PMID: 39620586 PMC: 11741935. DOI: 10.1111/imr.13426.

Helicase protein DDX11 as a novel antiviral factor promoting RIG-I-MAVS-mediated signaling pathway.

Zhang J, Zhang L, Liu D, Shi H, Zhang X, Chen J mBio. 2024; 15(12):e0202824.

PMID: 39470258 PMC: 11633105. DOI: 10.1128/mbio.02028-24.

Understanding the impact of transcription byproducts and contaminants.

Lenk R, Kleindienst W, Szabo G, Baiersdorfer M, Boros G, Keller J Front Mol Biosci. 2024; 11:1426129.

PMID: 39050733 PMC: 11266732. DOI: 10.3389/fmolb.2024.1426129.

A closer look at mammalian antiviral condensates.

Watkins J, Burke J Biochem Soc Trans. 2024; 52(3):1393-1404.

PMID: 38778761 PMC: 11234502. DOI: 10.1042/BST20231296.

Liquid-liquid phase separation in innate immunity.

Liu D, Yang J, Cristea I Trends Immunol. 2024; 45(6):454-469.

PMID: 38762334 PMC: 11247960. DOI: 10.1016/j.it.2024.04.009.

References

Gack M, Shin Y, Joo C, Urano T, Liang C, Sun L . TRIM25 RING-finger E3 ubiquitin ligase is essential for RIG-I-mediated antiviral activity. Nature. 2007; 446(7138):916-920. DOI: 10.1038/nature05732. View

Tran H, Schilling M, Wirbelauer C, Hess D, Nagamine Y . Facilitation of mRNA deadenylation and decay by the exosome-bound, DExH protein RHAU. Mol Cell. 2004; 13(1):101-11. DOI: 10.1016/s1097-2765(03)00481-7. View

Schmidt A, Schwerd T, Hamm W, Hellmuth J, Cui S, Wenzel M . 5'-triphosphate RNA requires base-paired structures to activate antiviral signaling via RIG-I. Proc Natl Acad Sci U S A. 2009; 106(29):12067-72. PMC: 2705279. DOI: 10.1073/pnas.0900971106. View

Sen A, Pruijssers A, Dermody T, Garcia-Sastre A, Greenberg H . The early interferon response to rotavirus is regulated by PKR and depends on MAVS/IPS-1, RIG-I, MDA-5, and IRF3. J Virol. 2011; 85(8):3717-32. PMC: 3126121. DOI: 10.1128/JVI.02634-10. View

Chalupnikova K, Lattmann S, Selak N, Iwamoto F, Fujiki Y, Nagamine Y . Recruitment of the RNA helicase RHAU to stress granules via a unique RNA-binding domain. J Biol Chem. 2008; 283(50):35186-98. PMC: 3259895. DOI: 10.1074/jbc.M804857200. View

Matsui T, Tanihara K, Date T . Expression of unphosphorylated form of human double-stranded RNA-activated protein kinase in Escherichia coli. Biochem Biophys Res Commun. 2001; 284(3):798-807. DOI: 10.1006/bbrc.2001.5039. View

Takahasi K, Yoneyama M, Nishihori T, Hirai R, Kumeta H, Narita R . Nonself RNA-sensing mechanism of RIG-I helicase and activation of antiviral immune responses. Mol Cell. 2008; 29(4):428-40. DOI: 10.1016/j.molcel.2007.11.028. View

Momose F, Kikuchi Y, Komase K, Morikawa Y . Visualization of microtubule-mediated transport of influenza viral progeny ribonucleoprotein. Microbes Infect. 2007; 9(12-13):1422-33. DOI: 10.1016/j.micinf.2007.07.007. View

Zhang Z, Kim T, Bao M, Facchinetti V, Jung S, Ghaffari A . DDX1, DDX21, and DHX36 helicases form a complex with the adaptor molecule TRIF to sense dsRNA in dendritic cells. Immunity. 2011; 34(6):866-78. PMC: 3652560. DOI: 10.1016/j.immuni.2011.03.027. View

10.

Gilfoy F, Mason P . West Nile virus-induced interferon production is mediated by the double-stranded RNA-dependent protein kinase PKR. J Virol. 2007; 81(20):11148-58. PMC: 2045557. DOI: 10.1128/JVI.00446-07. View

11.

Anderson P, Kedersha N . Stress granules: the Tao of RNA triage. Trends Biochem Sci. 2008; 33(3):141-50. DOI: 10.1016/j.tibs.2007.12.003. View

12.

Hornung V, Ellegast J, Kim S, Brzozka K, Jung A, Kato H . 5'-Triphosphate RNA is the ligand for RIG-I. Science. 2006; 314(5801):994-7. DOI: 10.1126/science.1132505. View

13.

Onomoto K, Jogi M, Yoo J, Narita R, Morimoto S, Takemura A . Critical role of an antiviral stress granule containing RIG-I and PKR in viral detection and innate immunity. PLoS One. 2012; 7(8):e43031. PMC: 3418241. DOI: 10.1371/journal.pone.0043031. View

14.

Pindel A, Sadler A . The role of protein kinase R in the interferon response. J Interferon Cytokine Res. 2010; 31(1):59-70. DOI: 10.1089/jir.2010.0099. View

15.

Fullam A, Schroder M . DExD/H-box RNA helicases as mediators of anti-viral innate immunity and essential host factors for viral replication. Biochim Biophys Acta. 2013; 1829(8):854-65. PMC: 7157912. DOI: 10.1016/j.bbagrm.2013.03.012. View

16.

Balachandran S, Barber G . PKR in innate immunity, cancer, and viral oncolysis. Methods Mol Biol. 2008; 383:277-301. DOI: 10.1007/978-1-59745-335-6_18. View

17.

Okonski K, Samuel C . Stress granule formation induced by measles virus is protein kinase PKR dependent and impaired by RNA adenosine deaminase ADAR1. J Virol. 2012; 87(2):756-66. PMC: 3554044. DOI: 10.1128/JVI.02270-12. View

18.

Booy E, Meier M, Okun N, Novakowski S, Xiong S, Stetefeld J . The RNA helicase RHAU (DHX36) unwinds a G4-quadruplex in human telomerase RNA and promotes the formation of the P1 helix template boundary. Nucleic Acids Res. 2012; 40(9):4110-24. PMC: 3351167. DOI: 10.1093/nar/gkr1306. View

19.

Creacy S, Routh E, Iwamoto F, Nagamine Y, Akman S, Vaughn J . G4 resolvase 1 binds both DNA and RNA tetramolecular quadruplex with high affinity and is the major source of tetramolecular quadruplex G4-DNA and G4-RNA resolving activity in HeLa cell lysates. J Biol Chem. 2008; 283(50):34626-34. PMC: 2596407. DOI: 10.1074/jbc.M806277200. View

20.

Kageyama M, Takahasi K, Narita R, Hirai R, Yoneyama M, Kato H . 55 Amino acid linker between helicase and carboxyl terminal domains of RIG-I functions as a critical repression domain and determines inter-domain conformation. Biochem Biophys Res Commun. 2011; 415(1):75-81. DOI: 10.1016/j.bbrc.2011.10.015. View