cis-Acting signals in encapsidation of Hantaan virus S-segment viral genomic RNA by its N protein.
The nucleocapsid (N) protein encapsidates both viral genomic RNA (vRNA) and the antigenomic RNA (cRNA), but not viral mRNA. Previous work has shown that the N protein has preference for vRNA, and this suggested the possibility of a cis-acting signal that could be used to initiate encapsidation for the S segment. To map the cis-acting determinants, several deletion RNA derivatives and synthetic oligoribonucleotides were constructed from the S segment of the Hantaan virus (HTNV) vRNA. N protein-RNA interactions were examined by UV cross-linking studies, filter-binding assays, and gel electrophoresis mobility shift assays to define the ability of each to bind HTNV N protein. The 5' end of the S-segment vRNA was observed to be necessary and sufficient for the binding reaction. Modeling of the 5' end of the vRNA revealed a possible stem-loop structure (SL) with a large single-stranded loop. We suggest that a specific interaction occurs between the N protein and sequences within this region to initiate encapsidation of the vRNAs.
Graduate Program in Molecular Biology, New Mexico State University, Las Cruces, New Mexico 88003, USA.
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Severson
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Xu
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Jonsson
D001483:Base Sequence; D001667:Binding, Competitive; D002213:Capsid / Q000378:metabolism*; D036022:Capsid Proteins; D015967:Gene Expression Regulation, Viral; D016679:Genome, Viral; D018063:Hantaan virus / Q000378:metabolism*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009843:Oligoribonucleotides / Q000138:chemical synthesis / Q000737:chemistry; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014758:Viral Core Proteins / Q000378:metabolism*
D036022:Capsid Proteins; D009843:Oligoribonucleotides; D012367:RNA, Viral; D014758:Viral Core Proteins; C111782:nucleocapsid protein, Hantaan virus