Information de reference pour ce titreAccession Number: | 00078401-201203000-00006.
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Author: | Zinzula, Luca; Esposito, Francesca; Pala, Daniela; Tramontano, Enzo *
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Institution: | Department of Life and Environmental Sciences, University of Cagliari, Cittadella di Monserrato SS554, 09042 Monserrato (Cagliari), Italy
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Title: | |
Source: | Antiviral Research. 93(3):354-363, March 2012.
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Abstract: | : The Ebola viruses (EBOVs) VP35 protein is a multifunctional major virulence factor involved in EBOVs replication and evasion of the host immune system. EBOV VP35 is an essential component of the viral RNA polymerase, it is a key participant of the nucleocapsid assembly and it inhibits the innate immune response by antagonizing RIG-I like receptors through its dsRNA binding function and, hence, by suppressing the host type I interferon (IFN) production. Insights into the VP35 dsRNA recognition have been recently revealed by structural and functional analysis performed on its C-terminus protein. We report the biochemical characterization of the Zaire ebolavirus (ZEBOV) full-length recombinant VP35 (rVP35)-dsRNA binding function. We established a novel in vitro magnetic dsRNA binding pull down assay, determined the rVP35 optimal dsRNA binding parameters, measured the rVP35 equilibrium dissociation constant for heterologous in vitro transcribed dsRNA of different length and short synthetic dsRNA of 8 bp, and validated the assay for compound screening by assessing the inhibitory ability of auryntricarboxylic acid (IC50 value of 50 [mu]g/mL). Furthermore, we compared the dsRNA binding properties of full length wt rVP35 with those of R305A, K309A and R312A rVP35 mutants, which were previously reported to be defective in dsRNA binding-mediated IFN inhibition, showing that the latter have measurably increased Kd values for dsRNA binding and modified migration patterns in mobility shift assays with respect to wt rVP35. Overall, these results provide the first characterization of the full-length wt and mutants VP35-dsRNA binding functions.
(C) 2012Elsevier, Inc.
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Author Keywords: | Ebola virus; VP35; dsRNA binding; Biochemical screening assay.
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References: | Barrette, R.W., Metwally, S.A., Rowland, J.M., Xu, L., Zaki, S.R., Nichol, S.T., Rollin, P.E., Towner, J.S., Shieh, W.-J., Batten, B., Sealy, T.K., Carrillo, C., Moran, K.E., Bracht, A.J., Mayr, G.A., Sirios-Cruz, M., Cathagan, D.P., Lautner, E.A., Ksiazek, T.G., White, W.R., McIntosh, M.T. 2009. Discovery of Swine as a host for the Reston ebolavirus. Science. 325, 204-206.
Barrette, R.W., Xu, L., Rowland, J.M., McIntosh, M.T., in press. Current perspectives on the phylogeny of Filoviridae. Infect. Genet. Evol. doi: 10.1016/j.meegid.2011.06.017
Basler, C.F., Wuang, X., Muhlberger, E., Volchkov, V., Paragas, J., Klenk, H.-D., Garcia-Sastre, A., Palese, P. 2000. The Ebola virus VP35 protein functions as a type I IFN antagonist. Proc. Natl. Acad. Sci. USA. 97, 12289-12294.
Basler, C.F., Mikulasova, A., Martinez-Sobrido, L., Paragas, J., Muhlberger, E., Bray, M., Klenk, H.-D., Palese, P., Garcia-Sastre, A. 2003. The Ebola virus VP35 protein inhibits activation of interferon regulatory factor 3. J. Virol. 77, 7945-7956.
Basler, C.F., Amarasinghe, G.K. 2009. Evasion of interferon responses by Ebola and Marburg viruses. J. Interferon Cytokine Res. 29, 511-520.
Becquart, P., Wauquier, N., Mahlakov, T., Nkoghe, D., Padilla, C., Souris, M., Ollomo, B., Gonzales, J.-P., De Lamballerie, X., Kazanji, M., Leroy, E.M. High prevalence of both humoral and cellular immunity to Zaire ebolavirus among rural populations in Gabon. PLoS One. 2010;5:e9126.
Boehmann, Y., Enterlein, S., Randolf, A., Muhlberger, E. 2005. A reconstituted replication and transcription system for Ebola virus Reston and comparison with Ebola virus Zaire. Virology. 332, 406-417.
Bowie, A.G., Unterholzner, L. 2008. Viral evasion and subversion of pattern-recognition receptor signaling. Nat. Rev. Immunol. 8, 911-922.
Bylund, D.B., Murrin, L.C. 2000. Radioligand saturation binding experiments over large concentration ranges. Life Sci. 67, 2897-2911.
Cardenas, W.B., Loo, Y.M., Gale, M. Jr, Hartman, A.L., Kimberlin, C.R., Martinez-Sobrido, L., Saphire, E.O., Basler, C.F. 2006. Ebola virus VP35 protein binds double-stranded RNA and inhibits alpha/beta interferon production induced by RIG-I signaling. J. Virol. 80, 5168-5178.
Chang, T.H., Kubota, T., Matsuoka, M., Jones, S., Bradfute, S.B., Bray, M., Ozato, K. Ebola Zaire virus blocks type I interferon production by exploiting the host SUMO modification machinery. PLoS Pathog. 2009;5:e1000493.
Chen, Y., Bopda-Waffo, A., Basu, A., Krishnan, R., Silberstein, E., Taylor, D.R., Talele, T.T., Arora, P., Kaushik-Basu, N. 2009. Characterization of aurintricarboxylic acid as a potent hepatitis C virus replicase inhibitor. Antivir. Chem. Chemother. 20, 19-36.
Chien, C., Xu, Y., Xiao, R., Aramini, J.M., Sahasrabudhe, P.V., Krug, R.M., Montelione, G.T. 2004. Biophysical characterization of the complex between double-stranded RNA and the N-terminal domain of the NS1 protein from influenza A virus: evidence for a novel RNA-binding mode. Biochemistry. 43, 1950-1962.
Chung, S., Wendeler, M., Rausch, J.W., Beilhartz, G., Gotte, M., O'Keefe, B.R., Bermingham, A., Beutler, J.A., Liu, S., Zhuang, X., Le Grice, S.F. 2010. Structure-activity analysis of vinylogous urea inhibitors of human immunodeficiency virus-encoded ribonuclease H. Antimicrob. Agents Chemother. 54, 3913-3921.
Conzelmann, K.K., 2005. Transcriptional activation of alpha/beta interferon genes: interference by non segmented negative-stranded RNA viruses. J. Virol. 79, 5241-5248.
Cushman, M., Sherman, P. 1992. Inhibition of HIV-1 integration protein by aurintricarboxylic acid monomers, monomer analogs, and polymer fractions. Biochim. Biophys. Res. Commun. 185, 85-90.
Enterlein, S., Warfield, K.L., Swenson, D.L., Stein, D.A., Smith, J.L., Gamble, C.S., Kroeker, A.D., Iversen, P.L., Bavari, S., Muhlberger, E. 2006. VP35 knockdown inhibits Ebola virus amplification and protects against lethal infection in mice. Antimicrob. Agents Chemother. 50, 984-993.
Fabozzi, G., Nabel, C.S., Dolan, M.A., Sullivan, N.J. 2011. Ebolavirus proteins suppress the effects of small interfering RNA by direct interaction with the mammalian RNA interference pathway. J. Virol. 85, 2512-2523.
Falzarano, D., Geisbert, T.W., Feldmann, H. 2011. Progress in filovirus vaccine development: evaluating the potential for clinical use. Expert. Rev. Vaccines. 10, 63-77.
Feldmann, H., Geisbert, T.W. 2011. Ebola hemorrhagic fever. Lancet. 377, 849-862.
Feng, Z., Cerveny, M., Yan, Z., He, B. 2007. The VP35 protein of Ebola virus inhibits the antiviral effect mediated by double-stranded RNA-dependent protein kinase PKR. J. Virol. 81, 182-192.
Geisbert, T.W., Lee, A.C.H., Robbins, M., Geisbert, J.B., Honko, A.N., Sood, V., Johnson, J.C., de Jong, S., Tavakoli, I., Judge, A., Hensley, L.E., Mac Lachlan, I. 2010. Post exposure protection of non-human primates against a lethal Ebola virus challenge with RNA interference: a proof of concept study. Lancet. 375, 1896-1905.
Ghosh, U., Giri, K., Bhattacharyya, N.P. 2009. Interaction of auryntricarboxylic acid (ATA) with four nucleic acid proteins DNase I, RNase A, reverse transcriptase and Taq polymerase. Spectrochim. Acta, Part A. 74, 1145-1151.
Gonzales, R.G., Haxo, R.S., Schleich, T. 1980. Mechanism of action of polymeric auryntricarboxylic acid, a potent inhibitor of protein-nucleic acid interactions. Biochemistry. 19, 4299-4303.
Haasnoot, J., de Vries, W., Geutjes, E.-J., Prins, M., de Haan, P., Berkhout, P. The Ebola virus VP35 protein is a suppressor of RNA silencing. PLoS Pathog. 2007;3:e86.
Hartman, A.L., Towner, J.S., Nichol, S.T. 2004. A C-terminal basic amino acid motif of Zaire ebolavirus VP35 is essential for type I interferon antagonism and displays high identity with the RNA-binding domain of another interferon antagonist, the NS1 protein of influenza A virus. Virology. 328, 177-184.
Hartman, A.L., Dover, J.E., Towner, J.S., Nichol, S.T. 2006. Reverse genetic generation of recombinant Zaire Ebola Viruses containing disrupted IRF-3 inhibitory domains results in attenuated virus growth in vitro and higher levels of IRF-3 activation without inhibiting viral transcription or replication. J. Virol. 80, 6430-6440.
Hartman, A.L., Bird, B.H., Towner, J.S., Antoniadou, Z.-A., Zaki, S.R., Nichol, S.T. 2008. Inhibition of IRF-3 activation by VP35 is critical for the high level of virulence of Ebola virus. J. Virol. 82, 2699-2704.
Hartman, A.L., Ling, L., Nichol, S.T., Hibberd, M.L. 2008. Whole-genome expression profiling reveals that inhibition of host innate immune response pathways by Ebola virus can be reversed by a single amino acid change in the VP35 protein. J. Virol. 82, 5348-5358.
Hartman, A.L., Towner, J.S., Nichol, S.T. 2010. Ebola and Marburg hemorrhagic fever. Clin. Lab. Med. 30, 161-177.
He, R., Adonov, A., Traykova-Adonova, M., Cao, J., Cutts, T., Grudesky, E., Deschambaul, Y., Berry, J., Drebot, M., Li, X. 2004. Potent and selective inhibition of SARS coronavirus replication by aurintricarboxylic acid. Biochem. Biophys. Res. Commun. 320, 1199-1203.
Huang, Y., Xu, L., Sun, Y., Nabel, G.J. 2002. The assembly of Ebola virus nucleocapsid requires virion-associated proteins 35 and 24 and posttranslational modification of nucleoprotein. Mol. Cell. 10, 307-316.
Jin, H., Yan, Z., Prabakhar, B.S., Feng, Z., Ma, Y., Verpooten, D., Ganesh, B., He, B. 2010. The VP35 protein of Ebola virus impairs dendritic cell maturation induced by virus and lipopolysaccharide. J. Gen. Virol. 91, 352-361.
John, S.P., Wang, T., Steffen, S., Longhi, S., Schmaljohn, C.S., Jonsson, C.B. 2007. VP30 is an RNA binding protein. J. Virol. 81, 8967-8976.
Johnson, R.F., McCarthy, S.E., Godlewski, P.J., Harty, R.N. 2006. Ebola virus VP35-VP40 interaction is sufficient for packaging 3E-5E minigenome RNA into virus-like particles. J. Virol. 80, 5135-5144.
Katze, M.G., Fornek, J.L., Palermo, R.E., Walters, K.-A., Korth, M.J. 2008. Innate immune modulation by RNA viruses: emerging insights from functional genomics. Nat. Rev. Immunol. 8, 644-654.
Kimberlin, C.R., Bornholdt, Z.A., Li, S., Woods, V.L. Jr, MacRae, I.J., Saphire, E.O. 2010. Ebolavirus VP35 uses a bimodal strategy to bind dsRNA for innate immune suppression. Proc. Natl. Acad. Sci. USA. 107, 314-319.
Kobinger, G.P., Leung, A., Neufeld, J., Richardson, J.S., Falzarano, D., Smith, G., Tierney, K., Patel, A., Weingartl, H.M. 2011. Replication, pathogenicity, shedding and transmission of Zaire ebolavirus in pigs. J. Infect. Dis. 204, 200-208.
Koyama, S., Ishii, K.J., Coban, C., Akira, S. 2008. Innate immune response to viral infection. Cytokine. 43, 336-341.
Kuhn, J.H., Becker, S., Ebihara, H., Geisbert, T.W., Johnson, K.M., Kawaoka, Y., Lipkin, W.I., Negredo, A.I., Netesov, S.V., Nichol, S.T., Palacios, G., Peters, C.J., Tenorio, A., Volchkov, V.E., Jahrling, P.B. 2010. Proposal for a revised taxonomy of the family Filoviridae: classification, names of taxa and viruses, and virus abbreviations. Arch. Virol. 155, 2083-2103.
Lemaire, P.A., Anderson, E., Lary, J., Cole, J.L. 2008. Mechanism of PKR activation by dsRNA. J. Mol. Biol. 381, 351-360.
Leroy, E.M., Gonzales, J.-P., Baize, S. 2011. Ebola and Marburg hemorrhagic fever viruses: major scientific advances, but a relatively minor public health threat for Africa. Clin. Microbiol. Infect. 17, 964-976.
Leung, D.W., Ginder, N.D., Fulton, D.B., Nix, J., Basler, C.F., Honzatko, R.B., Amarasinghe, G.K. 2009. Structure of the Ebola VP35 interferon inhibitory domain. Proc. Natl. Acad. Sci. USA. 106, 441-446.
Leung, D.W., Borek, D., Farahbakhsh, M., Ramanan, P., Nix, J.C., Wang, T., Prins, K.C., Otwinowski, Z., Honzatko, R.B., Helgeson, L.A., Basler, C.F., Amarasinghe, G.K. 2010. Crystallization and preliminary X-ray analysis of Ebola VP35 interferon inhibitory domain mutant proteins. Acta Crystallogr. Sect. F: Struct. Biol. Cryst. Commun. 66, 689-692.
Leung, D.W., Prins, K.C., Basler, C.F., Amarasinghe, G.K. 2010. Ebolavirus VP35 is a multifunctional virulence factor. Virulence. 6, 526-531.
Leung, D.W., Prins, K.C., Borek, D.M., Farahbakhsh, M., Tufariello, J.M., Ramaman, P., Nix, J.C., Helgeson, L.A., Otwinowski, Z., Honzatko, R.B., Basler, C.F., Amarasinghe, G.K. 2010. Structural basis for dsRNA recognition and interferon antagonism by Ebola VP35. Nat. Struct. Mol. Biol. 17, 165-172.
Leung, D.W., Shabman, R.S., Farahbakhsh, M., Prins, K.C., Borek, D.M., Wang, T., Muhlberger, E., Basler, C.F., Amarasinghe, G.K. 2010. Structural and functional characterization of Reston Ebola virus VP35 interferon inhibitory domain. J. Mol. Biol. 3, 347-357.
Loo, Y.-M., Gale, M. Jr. 2011. Immune signaling by RIG-I-like receptors. Immunity. 34, 680-692.
Lu, C., Ranjith-Kumar, C.T., Hao, L., ChengKao, C., Li, P. 2010. Crystal structure of RIG-I C-terminal domain bound to blunt-ended double-strand RNA without 5'triphosphate. Nucleic Acids Res. 39, 1565-1575.
Mahanty, S., Bray, M. 2004. Pathogenesis of filoviral hemorrhagic fevers. Lancet Infect. Dis. 4, 487-498.
Maroto, M., Fernandez, Y., Ortin, J., Pelaez, F., Cabello, M.A. 2008. Development of an HTS assay for the search of anti-influenza agents targeting the interaction of viral RNA with the NS1 protein. J. Biomol. Screen. 13, 581-590.
Mohamadzadeh, M., Chen, L., Schmaljohn, A.L. 2007. How Ebola and Marburg viruses battle the immune system. Nat. Rev. Immunol. 7, 556-567.
Mohamadzadeh, M., 2009. Potential factors induced by filoviruses that lead to immune suppression. Curr. Mol. Med. 9, 174-185.
Moller, P., Pariente, N., Klenk, H.-D., Becker, S. 2005. Homo-oligomerization of Marburgvirus VP35 is essential for its function in replication and transcription. J. Virol. 79, 14876-14886.
Motulsky, H.J., Neubig, R.R. 2010. Analyzing binding data. Curr. Protoc. Neurosci. 52, 751-7565. doi: 10.1002/0471142301.ns0705s52.
Muhlberger, E., Loftering, B., Klenk, H.-D., Becker, S. 1998. Three of the four nucleocapsid proteins of Marburg virus, NP, VP35, and L, are sufficient to mediate replication and transcription of Marburg virus-specific monocistronic minigenomes. J. Virol. 72, 8756-8764.
Muhlberger, E., Weik, M., Volchkov, V.E., Klenk, H.-D., Becker, S. 1999. Comparison of the transcription and replication strategies of Marburg virus and Ebola virus by using artificial replication systems. J. Virol. 73, 2333-2342.
Prins, K.C., Cardenas, W.B., Basler, C.F. 2009. Ebola virus protein VP35 impairs the function of interferon regulatory factor-activating kinases IKK epsilon and TBK-1. J. Virol. 83, 3069-3077.
Prins, K.C., Delpeut, S., Leung, D.W., Reynard, O., Volchkova, V.A., Reid, S.P., Ramanan, P., Cardenas, W.B., Amarasinghe, G.K., Volchkov, V.E., Basler, C.F. 2010. Mutations abrogating VP35 interactions with double-stranded RNA render Ebola virus avirulent in guinea pigs. J. Virol. 84, 3004-3015.
Randall, R.E., Goodbourn, S. 2008. Interferons and viruses: an interplay between induction, signaling, antiviral responses and virus countermeasures. J. Gen. Virol. 89, 1-47.
Reed, D.S., Lackemeyer, M.G., Garza, N.L., Sullivan, L.J., Nichols, D.K. 2011. Aerosol exposure to Zaire ebola virus in three non human primate species: differences in disease course and clinical pathology. Microbes Infect. 13, 930-936.
Reid, S.P., Cardenas, W.B., Basler, C.F. 2005. Homo-oligomerization facilitates the interferon-antagonist activity of the ebolavirus VP35 protein. Virology. 341, 179-189.
Sadler, A.J., Williams, B.R. 2008. Interferon-inducible antiviral effectors. Nat. Rev. Immunol. 8, 559-568.
Schumann, M., Gantke, T., Muhlberger, E. 2009. Ebola virus VP35 antagonizes PKR activity through its C-terminal interferon inhibitory domain. J. Virol. 83, 8993-8997.
Severson, W., Xu, X., Kuhn, M., Senutovitch, N., Thokala, M., Ferron, F., Longhi, S., Canard, B., Jonsson, C.B. 2005. Essential amino acids of the Hantaan virus N protein in its interaction with RNA. J. Virol. 79, 10032-10039.
Shi, W., Huang, Y., Sutton-Smith, M., Tissot, B., Panico, M., Morris, H.R., Dell, A., Haslam, S.M., Boyington, J., Graham, B.S., Yang, Z.-Y., Nabel, G.J. 2008. A filovirus-unique region of Ebola virus nucleoprotein confers aberrant migration and mediates its incorporation into virions. J. Virol. 82, 6190-6199.
Slice, L.W., Codner, E., Antelman, D., Holly, M., Wegrzynski, B., Wang, J., Toome, V., Hsu, M.C., Nalin, C.M. 1992. Characterization of recombinant HIV-1 Tat and its interaction with TAR RNA. Biochemistry. 31, 12062-12068.
Spiropoulou, C.F., Ranjan, P., Pearce, M.B., Sealy, T.K., Albarino, C.G., Gangappa, S., Fujita, T., Rollin, P.E., Nichol, S.T., Ksiazek, T.G., Sambhara, S. 2009. RIG-I activation inhibits ebola virus replication. Virology. 392, 11-15.
Tramontano, E., La Colla, P., Cheng, Y.-C. 1998. Biochemical characterization of the HIV-1 integrase 3'-processing activity and its inhibition by phosphorothioate oligonucleotides. Biochemistry. 37, 7237-7243.
Wang, Y., Ludwig, J., Schuberth, C., Goldeck, M., Schlee, M., Li, H., Juranek, S., Sheng, G., Micura, R., Tuschl, T., Hartmannand, G., Patel, D.J. 2010. Structural and functional insights into 5'-ppp RNA pattern recognition by the innate immune receptor RIG-I. Nat. Struct. Mol. Biol. 17, 781-787.
Warfield, K.L., Swenson, D.L., Olinger, G.G., Nichols, D.K., Pratt, W.D., Blouch, R., Stein, D.A., Aman, M.J., Iversen, P.I., Bavari, S. Gene-specific countermeasures against Ebola virus based on antisense phosphorodiamidate morpholino oligomers. PLoS Pathog. 2006;2:e1.
Wauquier, N., Becquart, P., Gasquet, C., Leroy, E.M. 2009. Immunoglobulin G in Ebola outbreak survivors. Gabon. Emerg. Infect. Dis. 15, 1136-1137.
Wauquier, N., Becquart, P., Padilla, C., Baize, S., Leroy, E.M. Human fatal Zaire Ebola virus infection is associated with an aberrant innate immunity and with massive lymphocyte apoptosis. PLoS Negl. Trop. Dis. 2010;5:e837.
Yap, Y., Zhang, X., Andonov, A., He, R. 2005. Structural analysis of inhibition mechanisms of aurintricarboxylic acid on SARS-CoV polymerase and other proteins. Comput. Biol. Chem. 29, 212-219.
Yoneyama, M., Fujita, T. 2010. Recognition of viral nucleic acids in innate immunity. Rev. Med. Virol. 20, 4-22.
Zampieri, C.A., Sullivan, N.J., Nabel, G.J. 2007. Immunopathology of highly virulent pathogens: insights from Ebola virus. Nat. Immunol. 8, 1159-1164.
Zheng, X., Bevilacqua, P.C. 2000. Straightening of bulged RNA by the double-stranded RNA-binding domain from the protein kinase PKR. Proc. Natl. Acad. Sci. USA. 97, 14162-14167.
Zinzula, L., Esposito, F., Muhlberger, E., Trunschke, M., Conrad, D., Piano, D., Tramontano, E. 2009. Purification and functional characterization of the full length recombinant Ebola virus VP35 protein expressed in E. coli. Protein Expr. Purif. 66, 113-119.
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Language: | English.
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Document Type: | Articles.
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Journal Subset: | Clinical Medicine. Pharmacology.
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ISSN: | 0166-3542
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DOI Number: | https://dx.doi.org/10.1016/j.ant...- ouverture dans une nouvelle fenêtre
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