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Journal of Virology, December 2007, p. 13754-13760, Vol. 81, No. 24
0022-538X/07/$08.00+0     doi:10.1128/JVI.01492-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

RNA Binding Domain of Jamestown Canyon Virus S Segment RNAs{triangledown}

Monica M. Ogg1,2 and Jean L. Patterson1*

Department of Virology and Immunology, Southwest Foundation for Biomedical Research, San Antonio, Texas 27227,1 Department of Microbiology and Immunology, The University of Texas Health Science Center at San Antonio, San Antonio, Texas 782292

Received 8 July 2007/ Accepted 21 September 2007

Jamestown Canyon virus (JCV) is a member of the Bunyaviridae family, Orthobunyavirus genus, California serogroup. Replication and, ultimately, assembly and packaging rely on the process of encapsidation. Therefore, the ability of viral RNAs (vRNAs) (genomic and antigenomic) to interact with the nucleocapsid protein (N protein) and the location of this binding domain on the RNAs are of interest. The questions to be addressed are the following. Where is the binding domain located on both the vRNA and cRNA strands, is this RNA bound when double or single stranded, and does this identified region have the ability to transform the binding potential of nonviral RNA? Full-length viral and complementary S segment RNA, as well as 3' deletion mutants of both vRNA and cRNA, nonviral RNA, and hybrid viral/nonviral RNA, were analyzed for their ability to interact with bacterially expressed JCV N protein. RNA-nucleocapsid interactions were examined by UV cross-linking, filter binding assays, and the generation of hybrid RNA to help define the area responsible for RNA-protein binding. The assays identified the region responsible for binding to the nucleocapsid as being contained within the 5' half of both the genomic and antigenomic RNAs. This region, if placed within nonviral RNA, is capable of altering the binding potential of nonviral RNA to levels seen with wild-type vRNAs.

* Corresponding author. Mailing address: Department of Virology and Immunology, P.O. Box 760549, San Antonio, TX 78245-0549. Phone: (210) 258-9431. Fax: (210) 670-3329. E-mail: jpatters{at}sfbr.org

{triangledown} Published ahead of print on 3 October 2007.

Journal of Virology, December 2007, p. 13754-13760, Vol. 81, No. 24
0022-538X/07/$08.00+0     doi:10.1128/JVI.01492-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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