Abstract
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.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Amino Acid Substitution
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Animals
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Aspartic Acid / genetics
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Aspartic Acid / metabolism
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Blotting, Western
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Carrier Proteins / genetics
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Carrier Proteins / metabolism
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Genes, Reporter
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Green Fluorescent Proteins / metabolism
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HEK293 Cells
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Humans
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Immunoprecipitation
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Interferon Type I / genetics
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Interferon Type I / metabolism
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Intracellular Signaling Peptides and Proteins
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Lymphocytic choriomeningitis virus / genetics
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Lymphocytic choriomeningitis virus / metabolism*
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Lymphocytic choriomeningitis virus / pathogenicity
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Lymphocytic choriomeningitis virus / physiology
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Molecular Sequence Data
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Mutation*
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Nucleoproteins / genetics
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Nucleoproteins / metabolism*
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Plasmids / genetics
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Plasmids / metabolism
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Protein Binding
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Protein Interaction Mapping
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RNA, Double-Stranded / genetics
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RNA, Double-Stranded / metabolism
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RNA, Viral / biosynthesis*
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RNA, Viral / metabolism
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RNA-Binding Proteins / genetics
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RNA-Binding Proteins / metabolism
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RNA-Dependent RNA Polymerase / genetics
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RNA-Dependent RNA Polymerase / metabolism
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Species Specificity
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Transcription, Genetic
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Transfection
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Viral Proteins / genetics
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Viral Proteins / metabolism*
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Virus Assembly*
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Virus Replication
Substances
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Carrier Proteins
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Interferon Type I
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Intracellular Signaling Peptides and Proteins
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Nucleoproteins
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RNA, Double-Stranded
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RNA, Viral
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RNA-Binding Proteins
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Viral Proteins
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p11 Z protein, Lymphocytic choriomeningitis virus
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Green Fluorescent Proteins
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Aspartic Acid
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RNA-Dependent RNA Polymerase