{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T11:44:37Z","timestamp":1790163877745,"version":"4.1.0"},"reference-count":62,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2012,10,16]],"date-time":"2012-10-16T00:00:00Z","timestamp":1350345600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Arenaviruses merit significant interest because several family members are etiological agents of severe hemorrhagic fevers, representing a major burden to public health. Currently, there are no FDA-licensed vaccines against arenaviruses and the only available antiviral therapy is limited to the use of ribavirin that is partially effective. Arenavirus nucleoprotein (NP) is found associated with the genomic RNA forming the viral ribonucleoproteins (vRNPs) that together with the polymerase (L) direct viral replication and transcription. Virion formation requires the recruitment of vRNPs into budding sites, a process in which the arenavirus matrix-like protein (Z) plays a major role. Therefore, proper NP-NP and NP-Z interactions are required for the generation of infectious progeny. In this work we demonstrate the role of the amino acid residue D471 in the self-association of lymphocytic choriomeningitis virus nucleoprotein (LCMV-NP). Amino acid substitutions at this position abrogate NP oligomerization, affecting its ability to mediate replication and transcription of a minigenome reporter plasmid. However, its ability to interact with the Z protein, counteract the cellular interferon response and bind to dsRNA analogs was retained. Additionally, we also document the dominant negative effect of D471G mutation on viral infection, suggesting that NP self-association is an excellent target for the development of new antivirals against arenaviruses.<\/jats:p>","DOI":"10.3390\/v4102137","type":"journal-article","created":{"date-parts":[[2012,10,16]],"date-time":"2012-10-16T11:36:59Z","timestamp":1350387419000},"page":"2137-2161","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["D471G Mutation in LCMV-NP Affects Its Ability to Self-associate and Results in a Dominant Negative Effect in Viral RNA Synthesis"],"prefix":"10.3390","volume":"4","author":[{"given":"Emilio","family":"Ortiz-Ria\u00f1o","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, University of Rochester, 601 Elmwood Avenue, Rochester, New York 14642, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benson","family":"Cheng","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Rochester, 601 Elmwood Avenue, Rochester, New York 14642, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan","family":"Torre","sequence":"additional","affiliation":[{"name":"Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California 92037, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7084-0804","authenticated-orcid":false,"given":"Luis","family":"Mart\u00ednez-Sobrido","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, University of Rochester, 601 Elmwood Avenue, Rochester, New York 14642, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2012,10,16]]},"reference":[{"key":"ref_1","first-page":"1792","article-title":"Arenaviridae: The viruses and their replication","volume":"Vol. 2","year":"2007","journal-title":"Fields virology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1006\/mpev.1997.0436","article-title":"Phylogenetic analysis of the Arenaviridae: patterns of virus evolution and evidence for cospeciation between arenaviruses and their rodent hosts","volume":"8","author":"Bowen","year":"1997","journal-title":"Mol Phylogenet. Evol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1007\/978-3-642-56029-3_4","article-title":"Lassa Fever","volume":"Vol. 262","year":"2002","journal-title":"Arenaviruses I"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1007\/978-3-642-56029-3_3","article-title":"Human infection with Arenaviruses in the Americas","volume":"Vol. 262","year":"2002","journal-title":"Arenaviruses I"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/S0166-3542(02)00202-4","article-title":"Arenaviruses other than Lassa virus","volume":"57","author":"Charrel","year":"2003","journal-title":"Antiviral. Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1080\/10408360490497456","article-title":"Lassa virus","volume":"41","author":"Gunther","year":"2004","journal-title":"Crit. Rev. Clin. Lab. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1707","DOI":"10.1086\/322620","article-title":"Viral hemorrhagic fever hazards for travelers in Africa","volume":"33","author":"Isaacson","year":"2001","journal-title":"Clin. Infect. Dis."},{"key":"ref_8","unstructured":"Seinbergas, M.M., Cibiriene, I.K., and Sverdlov Iu, M. (1976). [Main syndrome of prenatal infection caused by lymphocytic choriomeningitis virus - hydrocephalus and chorioretinal degeneration]. Pediatriia, 7\u201311."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1055\/s-0028-1107814","article-title":"[Prenatal infection with the virus of lymphocytic choriomeningitis: report of two cases (author's transl)]","volume":"99","author":"Ackermann","year":"1974","journal-title":"Dtsch. Med. Wochenschr."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2235","DOI":"10.1056\/NEJMoa053240","article-title":"Transmission of lymphocytic choriomeningitis virus by organ transplantation","volume":"354","author":"Fischer","year":"2006","journal-title":"N. Engl. J. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"991","DOI":"10.1056\/NEJMoa073785","article-title":"A new arenavirus in a cluster of fatal transplant-associated diseases","volume":"358","author":"Palacios","year":"2008","journal-title":"N. Engl. J. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"718","DOI":"10.1093\/clind\/24.4.718","article-title":"Treatment of Bolivian hemorrhagic fever with intravenous ribavirin","volume":"24","author":"Kilgore","year":"1997","journal-title":"Clin. Infect. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1056\/NEJM198601023140104","article-title":"Lassa fever. Effective therapy with ribavirin","volume":"314","author":"McCormick","year":"1986","journal-title":"N. Engl. J. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1304","DOI":"10.1128\/AAC.32.9.1304","article-title":"Ribavirin prophylaxis and therapy for experimental argentine hemorrhagic fever","volume":"32","author":"McKee","year":"1988","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1007\/978-3-642-56055-2_6","article-title":"Biology and pathogenesis of lymphocytic choriomeningitis virus infection","volume":"Vol. 263","year":"2002","journal-title":"Arenaviruses"},{"key":"ref_16","first-page":"1","article-title":"Lymphocytic choriomeningitis virus and immunology","volume":"263","author":"Zinkernagel","year":"2002","journal-title":"Curr. Top. Microbiol. Immunol."},{"key":"ref_17","first-page":"139","article-title":"Arenaviruses: genomic RNAs, transcription, and replication","volume":"262","author":"Meyer","year":"2002","journal-title":"Curr. Top Microbiol. Immunol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/j.virusres.2003.11.017","article-title":"Characterization of the Lassa virus matrix protein Z: electron microscopic study of virus-like particles and interaction with the nucleoprotein (NP)","volume":"100","author":"Eichler","year":"2004","journal-title":"Virus Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"3822","DOI":"10.1128\/JVI.79.6.3822-3830.2005","article-title":"Complementarity in the supramolecular design of arenaviruses and retroviruses revealed by electron cryomicroscopy and image analysis","volume":"79","author":"Neuman","year":"2005","journal-title":"J. Virol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"12978","DOI":"10.1073\/pnas.2133782100","article-title":"The small RING finger protein Z drives arenavirus budding: implications for antiviral strategies","volume":"100","author":"Perez","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","first-page":"E57","article-title":"Molecular and cell biology of the prototypic arenavirus LCMV: implications for understanding and combating hemorrhagic fever arenaviruses","volume":"1171 Suppl 1","year":"2009","journal-title":"Ann. N.Y. Acad. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"12701","DOI":"10.1073\/pnas.221447598","article-title":"The Lassa virus glycoprotein precursor GP-C is proteolytically processed by subtilase SKI-1\/S1P","volume":"98","author":"Lenz","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3882","DOI":"10.1128\/JVI.77.6.3882-3887.2003","article-title":"Role of the virus nucleoprotein in the regulation of lymphocytic choriomeningitis virus transcription and RNA replication","volume":"77","author":"Pinschewer","year":"2003","journal-title":"J. Virol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3470","DOI":"10.1128\/JVI.74.8.3470-3477.2000","article-title":"NP and L proteins of lymphocytic choriomeningitis virus (LCMV) are sufficient for efficient transcription and replication of LCMV genomic RNA analogs","volume":"74","author":"Lee","year":"2000","journal-title":"J. Virol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"9192","DOI":"10.1128\/JVI.00555-06","article-title":"Inhibition of the type I interferon response by the nucleoprotein of the prototypic arenavirus lymphocytic choriomeningitis virus","volume":"80","author":"Zuniga","year":"2006","journal-title":"J. Virol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"12696","DOI":"10.1128\/JVI.00882-07","article-title":"Differential inhibition of type I interferon induction by arenavirus nucleoproteins","volume":"81","author":"Giannakas","year":"2007","journal-title":"J. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"7728","DOI":"10.1128\/JVI.00187-12","article-title":"Arenavirus Nucleoprotein Targets Interferon Regulatory Factor-Activating Kinase IKK{varepsilon}","volume":"86","author":"Pythoud","year":"2012","journal-title":"J. Virol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"11330","DOI":"10.1128\/JVI.00763-09","article-title":"Identification of amino acid residues critical for the anti-interferon activity of the nucleoprotein of the prototypic arenavirus lymphocytic choriomeningitis virus","volume":"83","author":"Emonet","year":"2009","journal-title":"J. Virol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"779","DOI":"10.1038\/nature09605","article-title":"Cap binding and immune evasion revealed by Lassa nucleoprotein structure","volume":"468","author":"Qi","year":"2010","journal-title":"Nature"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2396","DOI":"10.1073\/pnas.1016404108","article-title":"Structure of the Lassa virus nucleoprotein reveals a dsRNA-specific 3' to 5' exonuclease activity essential for immune suppression","volume":"108","author":"Hastie","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"8185","DOI":"10.1128\/JVI.07240-11","article-title":"Arenavirus nucleoproteins prevent activation of nuclear factor kappa B","volume":"86","author":"Rodrigo","year":"2012","journal-title":"J. Virol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2443","DOI":"10.3390\/v2112443","article-title":"Inhibition of the type I interferon antiviral response during arenavirus infection","volume":"2","author":"Borrow","year":"2010","journal-title":"Viruses"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"13038","DOI":"10.1128\/JVI.05834-11","article-title":"The C-terminal region of lymphocytic choriomeningitis virus nucleoprotein contains distinct and segregable functional domains involved in NP-Z interaction and counteraction of the type I interferon response","volume":"85","author":"Cheng","year":"2011","journal-title":"J. Virol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3307","DOI":"10.1128\/JVI.05503-11","article-title":"Self-association of lymphocytic choriomeningitis virus nucleoprotein is mediated by its N-terminal region and is not required for its anti-interferon function","volume":"86","author":"Cheng","year":"2012","journal-title":"J. Virol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2012","DOI":"10.1128\/JVI.01875-10","article-title":"Identification of two functional domains within the arenavirus nucleoprotein","volume":"85","author":"Loureiro","year":"2011","journal-title":"J. Virol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"5415","DOI":"10.1128\/JVI.02417-09","article-title":"A role for the C terminus of Mopeia virus nucleoprotein in its incorporation into Z protein-induced virus-like particles","volume":"84","author":"Shtanko","year":"2010","journal-title":"J. Virol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"19365","DOI":"10.1073\/pnas.1108515108","article-title":"Crystal structure of the Lassa virus nucleoprotein-RNA complex reveals a gating mechanism for RNA binding","volume":"108","author":"Hastie","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"11443","DOI":"10.1128\/JVI.78.20.11443-11448.2004","article-title":"Myristoylation of the RING finger Z protein is essential for arenavirus budding","volume":"78","author":"Perez","year":"2004","journal-title":"J. Virol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"10700","DOI":"10.1128\/JVI.77.19.10700-10705.2003","article-title":"Lassa virus Z protein is a matrix protein and sufficient for the release of virus-like particles [corrected]","volume":"77","author":"Strecker","year":"2003","journal-title":"J. Virol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1186\/1743-422X-3-93","article-title":"The role of myristoylation in the membrane association of the Lassa virus matrix protein Z","volume":"3","author":"Strecker","year":"2006","journal-title":"Virol. J."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"6393","DOI":"10.1128\/JVI.76.12.6393-6397.2002","article-title":"Identification of the lymphocytic choriomeningitis virus (LCMV) proteins required to rescue LCMV RNA analogs into LCMV-like particles","volume":"76","author":"Lee","year":"2002","journal-title":"J. Virol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"4519","DOI":"10.1128\/JVI.79.7.4519-4526.2005","article-title":"Dual role of the lymphocytic choriomeningitis virus intergenic region in transcription termination and virus propagation","volume":"79","author":"Pinschewer","year":"2005","journal-title":"J. Virol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1684","DOI":"10.1128\/JVI.02229-10","article-title":"Use of single-cycle infectious lymphocytic choriomeningitis virus to study hemorrhagic fever arenaviruses","volume":"85","author":"Rodrigo","year":"2011","journal-title":"J. Virol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"5656","DOI":"10.1128\/JVI.00211-10","article-title":"Murine coronavirus delays expression of a subset of interferon-stimulated genes","volume":"84","author":"Rose","year":"2010","journal-title":"J. Virol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"11131","DOI":"10.1128\/JVI.05589-11","article-title":"Improved knockout methodology reveals that frog virus 3 mutants lacking either the 18K immediate-early gene or the truncated vIF-2alpha gene are defective for replication and growth in vivo","volume":"85","author":"Chen","year":"2011","journal-title":"J. Virol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"3473","DOI":"10.1073\/pnas.0900088106","article-title":"Generation of recombinant lymphocytic choriomeningitis viruses with trisegmented genomes stably expressing two additional genes of interest","volume":"106","author":"Emonet","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.virol.2008.09.008","article-title":"A cell-based luciferase assay amenable to high-throughput screening of inhibitors of arenavirus budding","volume":"382","author":"Capul","year":"2008","journal-title":"Virology"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1643","DOI":"10.1128\/JVI.75.4.1643-1655.2001","article-title":"Reverse genetics system for Uukuniemi virus (Bunyaviridae): RNA polymerase I-catalyzed expression of chimeric viral RNAs","volume":"75","author":"Flick","year":"2001","journal-title":"J. Virol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2157","DOI":"10.1128\/JVI.01433-09","article-title":"Hemagglutinin-pseudotyped green fluorescent protein-expressing influenza viruses for the detection of influenza virus neutralizing antibodies","volume":"84","author":"Cadagan","year":"2010","journal-title":"J. Virol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"664","DOI":"10.1002\/bit.22312","article-title":"A novel high-throughput cell-based method for integrated quantification of type I interferons and in vitro screening of immunostimulatory RNA drug delivery","volume":"103","author":"Nguyen","year":"2009","journal-title":"Biotechnol. Bioeng."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"191","DOI":"10.1016\/0166-0934(91)90018-U","article-title":"Quantification of lymphocytic choriomeningitis virus with an immunological focus assay in 24- or 96-well plates","volume":"33","author":"Battegay","year":"1991","journal-title":"J. Virol. Methods"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"360","DOI":"10.1126\/science.1125280","article-title":"Crystal structure of the rabies virus nucleoprotein-RNA complex","volume":"313","author":"Albertini","year":"2006","journal-title":"Science"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1279","DOI":"10.1126\/science.1177634","article-title":"Crystal structure of a nucleocapsid-like nucleoprotein-RNA complex of respiratory syncytial virus","volume":"326","author":"Tawar","year":"2009","journal-title":"Science"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"3638","DOI":"10.1096\/fj.08-112110","article-title":"Structure of the influenza virus A H5N1 nucleoprotein: implications for RNA binding, oligomerization, and vaccine design","volume":"22","author":"Ng","year":"2008","journal-title":"FASEB J."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1219","DOI":"10.1016\/j.str.2003.08.011","article-title":"Crystal structure of the borna disease virus nucleoprotein","volume":"11","author":"Rudolph","year":"2003","journal-title":"Structure"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1126\/science.1126953","article-title":"Structure of the vesicular stomatitis virus nucleoprotein-RNA complex","volume":"313","author":"Green","year":"2006","journal-title":"Science"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"W6","DOI":"10.1093\/nar\/gkm291","article-title":"Web services at the European bioinformatics institute","volume":"35","author":"Labarga","year":"2007","journal-title":"Nucleic Acids Res."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1184","DOI":"10.1128\/JVI.77.2.1184-1194.2003","article-title":"Characterization of the genomic promoter of the prototypic arenavirus lymphocytic choriomeningitis virus","volume":"77","author":"Perez","year":"2003","journal-title":"J. Virol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"2129","DOI":"10.1099\/0022-1317-72-9-2129","article-title":"Molecular organization of Junin virus S RNA: complete nucleotide sequence, relationship with other members of the Arenaviridae and unusual secondary structures","volume":"72 ( Pt 9)","author":"Ghiringhelli","year":"1991","journal-title":"J. Gen. Virol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"768","DOI":"10.1107\/S0907444902003359","article-title":"Conformations of amino acids in proteins","volume":"58","author":"Hovmoller","year":"2002","journal-title":"Acta Crystallogr. D. Biol. Crystallogr."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"12518","DOI":"10.1128\/JVI.05091-11","article-title":"Cross-species analysis of the replication complex of Old World arenaviruses reveals two nucleoprotein sites involved in L protein function","volume":"85","author":"Kerber","year":"2011","journal-title":"J. Virol."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"3743","DOI":"10.1128\/JVI.80.8.3743-3751.2006","article-title":"Functional mapping of the nucleoprotein of Ebola virus","volume":"80","author":"Watanabe","year":"2006","journal-title":"J. Virol."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/4\/10\/2137\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T21:52:52Z","timestamp":1760219572000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/4\/10\/2137"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,10,16]]},"references-count":62,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2012,10]]}},"alternative-id":["v4102137"],"URL":"https:\/\/doi.org\/10.3390\/v4102137","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2012,10,16]]}}}