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Journal of Virology, March 2001, p. 2646-2652, Vol. 75, No. 6
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.6.2646-2652.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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

William E. Severson,1 Xiaolin Xu,1 and Colleen B. Jonsson1,2,*

Graduate Program in Molecular Biology1 and Department of Chemistry and Biochemistry,2 New Mexico State University, Las Cruces, New Mexico

Received 17 October 2000/Accepted 16 December 2000

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.

* Corresponding author. Mailing address: Department of Chemistry and Biochemistry, MSC 3C, P.O. Box 30001, New Mexico State University, Las Cruces, NM 88003. Phone: (505) 646-3346. Fax: (505) 646-2649. E-mail: cjonsson{at}nmsu.edu.

Journal of Virology, March 2001, p. 2646-2652, Vol. 75, No. 6
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.6.2646-2652.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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