cis -Acting Signals in Encapsidation of Hantaan Virus S-Segment Viral Genomic RNA by Its N Protein

Author:

Severson William E.1,Xu Xiaolin1,Jonsson Colleen B.12

Affiliation:

1. Graduate Program in Molecular Biology1 and

2. Department of Chemistry and Biochemistry,2 New Mexico State University, Las Cruces, New Mexico

Abstract

ABSTRACT 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.

Publisher

American Society for Microbiology

Subject

Virology,Insect Science,Immunology,Microbiology

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