Interaction between molecules of hantavirus nucleocapsid protein.
Intermolecular interactions of Tula hantavirus N (nucleocapsid) protein were detected in the yeast two-hybrid system, prompting further attempts to study this phenomenon. Using chemical cross-linking and immunoblotting it was shown that the N protein from purified virus and from infected cell lysates as well as recombinant protein produced in a baculovirus expression system are capable of forming dimers, trimers and multimers, thus confirming the capacity of the protein molecules to interact with each other. An ELISA format was developed in which molecules of the recombinant N protein were shown to associate non-covalently, via electrostatic interactions. Divalent cations (Ca(2+), Mn(2+), Mg(2+), Ba(2+)) enhanced the process 3- to 8-fold suggesting that adequate folding of the N protein is crucial for the association. Based on these data a model for hantavirus nucleocapsid assembly is proposed, in which N molecules first trimerize around the viral RNA molecule, and then the trimers gradually assemble forming longer multimers.
Kaukinen
Pasi
P
Department of Virology, Haartman Institute, PO Box 21, FIN-00014 University of Helsinki, Finland1.
Koistinen
Vesa
V
Department of Virology, Haartman Institute, PO Box 21, FIN-00014 University of Helsinki, Finland1.
Vapalahti
Olli
O
Department of Virology, Haartman Institute, PO Box 21, FIN-00014 University of Helsinki, Finland1.
Vaheri
Antti
A
Department of Virology, Haartman Institute, PO Box 21, FIN-00014 University of Helsinki, Finland1.
Plyusnin
Alexander
A
Department of Virology, Haartman Institute, PO Box 21, FIN-00014 University of Helsinki, Finland1.
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001464:Barium / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D002213:Capsid / Q000378:metabolism; D036022:Capsid Proteins; D002413:Cations, Divalent; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D006476:Orthohantavirus / Q000378:metabolism*; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017510:Protein Folding; D011994:Recombinant Proteins / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D014758:Viral Core Proteins / Q000378:metabolism; D019065:Virus Assembly
D036022:Capsid Proteins; D002413:Cations, Divalent; D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins; D014758:Viral Core Proteins; C111782:nucleocapsid protein, Hantaan virus; D001464:Barium; D008345:Manganese; D008274:Magnesium; D002118:Calcium