Year-end Sale % OFF 
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.
Highlights
Hantaviruses are tripartite negative-sense RNA viruses in the Bunyaviridae family [1]
The proposed activities of the hantavirus N protein in the virus life cycle rely on differential interactions with the three types of viral RNAs, the vRNA, the virus complementary and the messenger RNA
To begin to address the biochemical basis of N protein and viral RNA interactions, we investigated the first event in the assembly process, the binding step
Summary
N, nucleocapsid; SNV, Sin Nombre virus; HTNV, Hantaan virus; PUUV, Puumala virus; DTT, dithiothreitol; VSV, vesicular stomatitis virus; PAGE, polyacrylamide gel electrophoresis; S ORF, S segment open reading frame; GEMSA, gel electrophoretic mobility shift assay. While vRNA and cRNA can form noncovalently, closed circular structures, mRNA cannot [13] This observation led to the suggestion that this gene region may play a role in assembly of the nucleocapsids, no sequences or secondary structures in any of the viral RNAs have been defined [14]. No readily identifiable RNA binding motifs have been located in N proteins, deletion mapping of Hantaan (HTNV) and Puumala virus (PUUV) N proteins identified a nonspecific RNA binding domain in the carboxyl-terminal 93 amino acids [15]. This region showed no apparent specificity for viral RNA. We explored the ability of a heterologous N protein from Sin Nombre virus (SNV) to interact with HTNV-derived RNAs and a nonviral RNA
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
