Cytoplasmic tails of hantavirus glycoproteins interact with the nucleocapsid protein.
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.
Hepojoki
J
J
Department of Virology, Haartman Institute, University of Helsinki, Finland. jussi.hepojoki@helsinki.fi
Strandin
T
T
Wang
H
H
Vapalahti
O
O
Vaheri
A
A
Lankinen
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D001665:Binding Sites / Q000235:genetics; D002522:Chlorocebus aethiops; D006476:Orthohantavirus / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D010449:Peptide Mapping; D054730:Protein Interaction Domains and Motifs; D025941:Protein Interaction Mapping; D029262:Puumala virus / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012261:Ribonucleoproteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D019590:Nucleocapsid Proteins; D012261:Ribonucleoproteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C100886:nucleocapsid protein, Puumala virus