Abstract
Here we characterize the interaction between the glycoproteins (Gn and Gc) and the ribonucleoprotein (RNP) of Puumala virus (PUUV; genus Hantavirus, family Bunyaviridae). The interaction was initially established with native proteins by co-immunoprecipitating PUUV nucleocapsid (N) protein with the glycoprotein complex. Mapping of the interaction sites revealed that the N protein has multiple binding sites in the cytoplasmic tail (CT) of Gn and is also able to bind to the predicted CT of Gc. The importance of Gn- and Gc-CTs to the recognition of RNP was further verified in pull-down assays using soluble peptides with binding capacity to both recombinant N protein and the RNPs of PUUV and Tula virus. Additionally, the N protein of PUUV was demonstrated to interact with peptides of Gn and Gc from a variety of hantavirus species, suggesting a conserved RNP-recognition mechanism within the genus. Based on these and our previous results, we suggest that the complete hetero-oligomeric (Gn-Gc)(4) spike complex of hantaviruses mediates the packaging of RNP into virions.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Antibodies, Monoclonal
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Antibodies, Neutralizing
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Binding Sites / genetics
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Chlorocebus aethiops
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Models, Molecular
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Molecular Sequence Data
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Nucleocapsid Proteins / chemistry*
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Nucleocapsid Proteins / genetics
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Nucleocapsid Proteins / immunology
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Orthohantavirus / chemistry*
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Orthohantavirus / genetics
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Orthohantavirus / immunology
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Peptide Mapping
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Protein Interaction Domains and Motifs
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Protein Interaction Mapping
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Puumala virus / chemistry
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Puumala virus / genetics
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Puumala virus / immunology
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Ribonucleoproteins / chemistry
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Ribonucleoproteins / genetics
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Ribonucleoproteins / immunology
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Sequence Homology, Amino Acid
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Vero Cells
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Viral Envelope Proteins / chemistry
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Viral Envelope Proteins / genetics
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Viral Envelope Proteins / immunology
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Viral Proteins / chemistry*
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Viral Proteins / genetics
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Viral Proteins / immunology
Substances
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Antibodies, Monoclonal
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Antibodies, Neutralizing
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Nucleocapsid Proteins
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Ribonucleoproteins
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Viral Envelope Proteins
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Viral Proteins
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nucleocapsid protein, Puumala virus