SARS-CoV-2 Nonstructural Proteins 1 and 13 Suppress Caspase-1 and the NLRP3 Inflammasome Activation
Abstract
:1. Introduction
2. Results
2.1. Screening of SARS-CoV-2 cDNA Libraries to Identify Viral ORFs That Inhibit Caspase-1 Activity
2.2. The Amino Acid Residues of NSP1 Critical for Host Translation Shutoff Are Required for Caspase-1 Inhibition
2.3. Both the N- and C-Termini of NSP13 Protein Are Required for Caspase-1 Inhibition
2.4. NSP1 and NSP13 Inhibit Activation of the NLRP3 Inflammasome in THP-1 Cells
3. Discussion
4. Materials and Methods
4.1. Cells, Reagents and Transfection
4.2. SARS-CoV-2 cDNA Library
4.3. Plasmid Constructs
4.4. Western Blot Analysis
4.5. Electroporation
4.6. Enzyme-linked Immunosorbent Assay (ELISA)
4.7. Caspase Glo®1 inflammasome Assay
4.8. Quantitative Reverse Transcription PCR (qRT-PCR)
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Du Toit, A. Outbreak of a novel coronavirus. Nat. Rev. Microbiol. 2020, 18, 123. [Google Scholar] [CrossRef]
- Sawicki, S.G.; Sawicki, D.L.; Siddell, S.G. A contemporary view of coronavirus transcription. J. Virol. 2007, 81, 20–29. [Google Scholar] [CrossRef] [Green Version]
- Guo, H.; Callaway, J.B.; Ting, J.P. Inflammasomes: Mechanism of action, role in disease, and therapeutics. Nat. Med. 2015, 21, 677–687. [Google Scholar] [CrossRef] [Green Version]
- Chung, W.C.; Kang, H.R.; Yoon, H.; Kang, S.J.; Ting, J.P.; Song, M.J. Influenza A virus NS1 protein inhibits the NLRP3 inflammasome. PLoS ONE 2015, 10, e0126456. [Google Scholar] [CrossRef]
- Gerlic, M.; Faustin, B.; Postigo, A.; Yu, E.C.; Proell, M.; Gombosuren, N.; Krajewska, M.; Flynn, R.; Croft, M.; Way, M.; et al. Vaccinia virus F1L protein promotes virulence by inhibiting inflammasome activation. Proc. Natl. Acad. Sci. USA 2013, 110, 7808–7813. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Gregory, S.M.; Davis, B.K.; West, J.A.; Taxman, D.J.; Matsuzawa, S.; Reed, J.C.; Ting, J.P.; Damania, B. Discovery of a viral NLR homolog that inhibits the inflammasome. Science 2011, 331, 330–334. [Google Scholar] [CrossRef]
- Haneklaus, M.; Gerlic, M.; Kurowska-Stolarska, M.; Rainey, A.A.; Pich, D.; McInnes, I.B.; Hammerschmidt, W.; O’Neill, L.A.; Masters, S.L. Cutting edge: miR-223 and EBV miR-BART15 regulate the NLRP3 inflammasome and IL-1beta production. J. Immunol. 2012, 189, 3795–3799. [Google Scholar] [CrossRef] [Green Version]
- Karim, R.; Meyers, C.; Backendorf, C.; Ludigs, K.; Offringa, R.; van Ommen, G.J.; Melief, C.J.; van der Burg, S.H.; Boer, J.M. Human papillomavirus deregulates the response of a cellular network comprising of chemotactic and proinflammatory genes. PLoS ONE 2011, 6, e17848. [Google Scholar] [CrossRef] [Green Version]
- Kettle, S.; Alcami, A.; Khanna, A.; Ehret, R.; Jassoy, C.; Smith, G.L. Vaccinia virus serpin B13R (SPI-2) inhibits interleukin-1beta-converting enzyme and protects virus-infected cells from TNF- and Fas-mediated apoptosis, but does not prevent IL-1beta-induced fever. J. Gen. Virol. 1997, 78, 677–685. [Google Scholar] [CrossRef]
- Komatsu, T.; Tanaka, Y.; Kitagawa, Y.; Koide, N.; Naiki, Y.; Morita, N.; Gotoh, B.; Yokochi, T. Sendai virus V protein inhibits the secretion of Interleukin-1β by preventing NLRP3 inflammasome assembly. J. Virol. 2018, 92. [Google Scholar] [CrossRef] [Green Version]
- Komune, N.; Ichinohe, T.; Ito, M.; Yanagi, Y. Measles virus V protein inhibits NLRP3 inflammasome-mediated interleukin-1β secretion. J. Virol. 2011, 85, 13019–13026. [Google Scholar] [CrossRef] [Green Version]
- Marshall, B.; Puthalakath, H.; Caria, S.; Chugh, S.; Doerflinger, M.; Colman, P.M.; Kvansakul, M. Variola virus F1L is a Bcl-2-like protein that unlike its vaccinia virus counterpart inhibits apoptosis independent of Bim. Cell Death Dis. 2015, 6, e1680. [Google Scholar] [CrossRef]
- Moriyama, M.; Chen, I.Y.; Kawaguchi, A.; Koshiba, T.; Nagata, K.; Takeyama, H.; Hasegawa, H.; Ichinohe, T. The RNA- and TRIM25-binding domains of Influenza virus NS1 protein are essential for suppression of NLRP3 inflammasome-mediated interleukin-1β secretion. J. Virol. 2016, 90, 4105–4114. [Google Scholar] [CrossRef] [Green Version]
- Niebler, M.; Qian, X.; Hofler, D.; Kogosov, V.; Kaewprag, J.; Kaufmann, A.M.; Ly, R.; Bohmer, G.; Zawatzky, R.; Rosl, F.; et al. Post-translational control of IL-1beta via the human papillomavirus type 16 E6 oncoprotein: A novel mechanism of innate immune escape mediated by the E3-ubiquitin ligase E6-AP and p53. PLoS Pathog. 2013, 9, e1003536. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Petit, F.; Bertagnoli, S.; Gelfi, J.; Fassy, F.; Boucraut-Baralon, C.; Milon, A. Characterization of a myxoma virus-encoded serpin-like protein with activity against interleukin-1 beta-converting enzyme. J. Virol. 1996, 70, 5860–5866. [Google Scholar] [CrossRef] [Green Version]
- Ray, C.A.; Black, R.A.; Kronheim, S.R.; Greenstreet, T.A.; Sleath, P.R.; Salvesen, G.S.; Pickup, D.J. Viral inhibition of inflammation: Cowpox virus encodes an inhibitor of the interleukin-1 beta converting enzyme. Cell 1992, 69, 597–604. [Google Scholar] [CrossRef]
- Shil, N.K.; Pokharel, S.M.; Banerjee, A.K.; Hoffman, M.; Bose, S. Inflammasome antagonism by human parainfluenza virus type 3C protein. J. Virol. 2018, 92. [Google Scholar] [CrossRef] [Green Version]
- Wang, H.; Lei, X.; Xiao, X.; Yang, C.; Lu, W.; Huang, Z.; Leng, Q.; Jin, Q.; He, B.; Meng, G.; et al. Reciprocal regulation between Enterovirus 71 and the NLRP3 inflammasome. Cell Rep. 2015, 12, 42–48. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Chan, Y.K.; Gack, M.U. Viral evasion of intracellular DNA and RNA sensing. Nat. Rev. Microbiol. 2016, 14, 360–373. [Google Scholar] [CrossRef]
- Stewart, M.K.; Cookson, B.T. Evasion and interference: Intracellular pathogens modulate caspase-dependent inflammatory responses. Nat. Rev. Microbiol. 2016, 14, 346–359. [Google Scholar] [CrossRef] [PubMed]
- Chen, I.Y.; Moriyama, M.; Chang, M.F.; Ichinohe, T. Severe acute respiratory syndrome coronavirus viroporin 3a activates the NLRP3 inflammasome. Front. Microbiol. 2019, 10, 50. [Google Scholar] [CrossRef] [Green Version]
- Nieto-Torres, J.L.; Verdia-Baguena, C.; Jimenez-Guardeno, J.M.; Regla-Nava, J.A.; Castano-Rodriguez, C.; Fernandez-Delgado, R.; Torres, J.; Aguilella, V.M.; Enjuanes, L. Severe acute respiratory syndrome coronavirus E protein transports calcium ions and activates the NLRP3 inflammasome. Virology 2015, 485, 330–339. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Shi, C.S.; Nabar, N.R.; Huang, N.N.; Kehrl, J.H. SARS-coronavirus open reading frame-8b triggers intracellular stress pathways and activates NLRP3 inflammasomes. Cell Death Discov. 2019, 5, 101. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Siu, K.L.; Yuen, K.S.; Castano-Rodriguez, C.; Ye, Z.W.; Yeung, M.L.; Fung, S.Y.; Yuan, S.; Chan, C.P.; Yuen, K.Y.; Enjuanes, L.; et al. Severe acute respiratory syndrome coronavirus ORF3a protein activates the NLRP3 inflammasome by promoting TRAF3-dependent ubiquitination of ASC. FASEB J. 2019, 33, 8865–8877. [Google Scholar] [CrossRef]
- Freeman, T.L.; Swartz, T.H. Targeting the NLRP3 Inflammasome in Severe COVID-19. Front. Immunol. 2020, 11, 1518. [Google Scholar] [CrossRef]
- Lee, S.; Channappanavar, R.; Kanneganti, T.D. Coronaviruses: Innate immunity, inflammasome activation, inflammatory cell death, and cytokines. Trends Immunol. 2020, 41, 1083–1099. [Google Scholar] [CrossRef] [PubMed]
- van den Berg, D.F.; Te Velde, A.A. Severe COVID-19: NLRP3 inflammasome dysregulated. Front. Immunol. 2020, 11, 1580. [Google Scholar] [CrossRef]
- Yap, J.K.Y.; Moriyama, M.; Iwasaki, A. Inflammasomes and pyroptosis as therapeutic targets for COVID-19. J. Immunol. 2020, 205, 307–312. [Google Scholar] [CrossRef] [PubMed]
- Huang, C.; Lokugamage, K.G.; Rozovics, J.M.; Narayanan, K.; Semler, B.L.; Makino, S. SARS coronavirus NSP1 protein induces template-dependent endonucleolytic cleavage of mRNAs: Viral mRNAs are resistant to nsp1-induced RNA cleavage. PLoS Pathog. 2011, 7, e1002433. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kamitani, W.; Narayanan, K.; Huang, C.; Lokugamage, K.; Ikegami, T.; Ito, N.; Kubo, H.; Makino, S. Severe acute respiratory syndrome coronavirus nsp1 protein suppresses host gene expression by promoting host mRNA degradation. Proc. Natl. Acad. Sci. USA 2006, 103, 12885–12890. [Google Scholar] [CrossRef] [Green Version]
- Narayanan, K.; Huang, C.; Lokugamage, K.; Kamitani, W.; Ikegami, T.; Tseng, C.T.; Makino, S. Severe acute respiratory syndrome coronavirus nsp1 suppresses host gene expression, including that of type I interferon, in infected cells. J. Virol. 2008, 82, 4471–4479. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wathelet, M.G.; Orr, M.; Frieman, M.B.; Baric, R.S. Severe acute respiratory syndrome coronavirus evades antiviral signaling: Role of nsp1 and rational design of an attenuated strain. J. Virol. 2007, 81, 11620–11633. [Google Scholar] [CrossRef] [Green Version]
- Lokugamage, K.G.; Narayanan, K.; Huang, C.; Makino, S. Severe acute respiratory syndrome coronavirus protein nsp1 is a novel eukaryotic translation inhibitor that represses multiple steps of translation initiation. J. Virol. 2012, 86, 13598–13608. [Google Scholar] [CrossRef] [Green Version]
- Tanaka, T.; Kamitani, W.; DeDiego, M.L.; Enjuanes, L.; Matsuura, Y. Severe acute respiratory syndrome coronavirus nsp1 facilitates efficient propagation in cells through a specific translational shutoff of host mRNA. J. Virol. 2012, 86, 11128–11137. [Google Scholar] [CrossRef] [Green Version]
- Zheng, D.; Liwinski, T.; Elinav, E. Inflammasome activation and regulation: Toward a better understanding of complex mechanisms. Cell Discov. 2020, 6, 36. [Google Scholar] [CrossRef] [PubMed]
- Chan, J.F.; Lau, S.K.; To, K.K.; Cheng, V.C.; Woo, P.C.; Yuen, K.Y. Middle East respiratory syndrome coronavirus: Another zoonotic betacoronavirus causing SARS-like disease. Clin. Microbiol. Rev. 2015, 28, 465–522. [Google Scholar] [CrossRef] [Green Version]
- Chen, J.; Subbarao, K. The Immunobiology of SARS. Annu. Rev. Immunol. 2007, 25, 443–472. [Google Scholar] [CrossRef]
- Tang, Y.; Liu, J.; Zhang, D.; Xu, Z.; Ji, J.; Wen, C. Cytokine storm in COVID-19: The current evidence and treatment strategies. Front. Immunol. 2020, 11, 1708. [Google Scholar] [CrossRef]
- Ahn, M.; Anderson, D.E.; Zhang, Q.; Tan, C.W.; Lim, B.L.; Luko, K.; Wen, M.; Chia, W.N.; Mani, S.; Wang, L.C.; et al. Dampened NLRP3-mediated inflammation in bats and implications for a special viral reservoir host. Nat. Microbiol. 2019, 4, 789–799. [Google Scholar] [CrossRef] [PubMed]
- Mirza, M.U.; Froeyen, M. Structural elucidation of SARS-CoV-2 vital proteins: Computational methods reveal potential drug candidates against main protease, Nsp12 polymerase and Nsp13 helicase. J. Pharm. Anal. 2020, 10, 320–328. [Google Scholar] [CrossRef]
- Seybert, A.; Posthuma, C.C.; van Dinten, L.C.; Snijder, E.J.; Gorbalenya, A.E.; Ziebuhr, J. A complex zinc finger controls the enzymatic activities of nidovirus helicases. J. Virol. 2005, 79, 696–704. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Tate, M.D.; Ong, J.D.H.; Dowling, J.K.; McAuley, J.L.; Robertson, A.B.; Latz, E.; Drummond, G.R.; Cooper, M.A.; Hertzog, P.J.; Mansell, A. Reassessing the role of the NLRP3 inflammasome during pathogenic influenza A virus infection via temporal inhibition. Sci. Rep. 2016, 6, 27912. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kang, S.C.; Lim, S.Y.; Song, Y.J. Lupeol is one of active components in the extract of Chrysanthemum indicum Linne that inhibits LMP1-induced NF-κB activation. PLoS ONE 2013, 8, e82688. [Google Scholar] [CrossRef]
- Kim, D.K.; Knapp, J.J.; Kuang, D.; Chawla, A.; Cassonnet, P.; Lee, H.; Sheykhkarimli, D.; Samavarchi-Tehrani, P.; Abdouni, H.; Rayhan, A.; et al. A Comprehensive, Flexible Collection of SARS-CoV-2 Coding Regions. G3 Genes Genomes Genet. 2020, 10, 3399–3402. [Google Scholar] [CrossRef] [PubMed]
- Kim, J.E.; Kim, S.Y.; Lim, S.Y.; Kieff, E.; Song, Y.J. Role of Ca2+/calmodulin-dependent kinase II-IRAK1 interaction in LMP1-induced NF-kappaB activation. Mol. Cell Biol. 2014, 34, 325–334. [Google Scholar] [CrossRef] [PubMed] [Green Version]
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Kim, N.-E.; Kim, D.-K.; Song, Y.-J. SARS-CoV-2 Nonstructural Proteins 1 and 13 Suppress Caspase-1 and the NLRP3 Inflammasome Activation. Microorganisms 2021, 9, 494. https://doi.org/10.3390/microorganisms9030494
Kim N-E, Kim D-K, Song Y-J. SARS-CoV-2 Nonstructural Proteins 1 and 13 Suppress Caspase-1 and the NLRP3 Inflammasome Activation. Microorganisms. 2021; 9(3):494. https://doi.org/10.3390/microorganisms9030494
Chicago/Turabian StyleKim, Na-Eun, Dae-Kyum Kim, and Yoon-Jae Song. 2021. "SARS-CoV-2 Nonstructural Proteins 1 and 13 Suppress Caspase-1 and the NLRP3 Inflammasome Activation" Microorganisms 9, no. 3: 494. https://doi.org/10.3390/microorganisms9030494