title: Characterization of the Hantaan nucleocapsid protein-ribonucleic acid interaction.
abstract: The nucleocapsid (N) protein functions in hantavirus replication through its interactions with the viral genomic and antigenomic RNAs. To address the biological functions of the N protein, it was critical to first define this binding interaction. The dissociation constant, K(d), for the interaction of the Hantaan virus (HTNV) N protein and its genomic S segment (vRNA) was measured under several solution conditions. Overall, increasing the NaCl and Mg(2+) in these binding reactions had little impact on the K(d). However, the HTNV N protein showed an enhanced specificity for HTNV vRNA as compared with the S segment open reading frame RNA or a nonviral RNA with increasing ionic strength and the presence of Mg(2+). In contrast, the assembly of Sin Nombre virus N protein-HTNV vRNA complexes was inhibited by the presence of Mg(2+) or an increase in the ionic strength. The K(d) values for HTNV and Sin Nombre virus N proteins were nearly identical for the S segment open reading frame RNA, showing weak affinity over several binding reaction conditions. Our data suggest a model in which specific recognition of the HTNV vRNA by the HTNV N protein resides in the noncoding regions of the HTNV vRNA.
authors.affiliation: Graduate Program in Molecular Biology, New Mexico State University, Las Cruces, New Mexico 88003, USA.
authors.forename: W
authors.initials: W
authors.lastname: Severson
authors.forename: L
authors.initials: L
authors.lastname: Partin
authors.forename: C S
authors.initials: CS
authors.lastname: Schmaljohn
authors.forename: C B
authors.initials: CB
authors.lastname: Jonsson
mesh_terms: D002213:Capsid / Q000302:isolation & purification / Q000378:metabolism*; D036022:Capsid Proteins; D018063:Hantaan virus / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism; D009994:Osmolar Concentration; D011485:Protein Binding; D012367:RNA, Viral / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D014758:Viral Core Proteins / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D036022:Capsid Proteins; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D014758:Viral Core Proteins; C111782:nucleocapsid protein, Hantaan virus; D008274:Magnesium