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Journal of Virology, October 2005, p. 13166-13172, Vol. 79, No. 20
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.20.13166-13172.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Homotypic Interaction of Bunyamwera Virus Nucleocapsid Protein

Vincent H. J. Leonard,{dagger} Alain Kohl, Jane C. Osborne,{ddagger} Angela McLees, and Richard M. Elliott*

Division of Virology, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G11 5JR, Scotland, United Kingdom

Received 12 May 2005/ Accepted 19 July 2005

The bunyavirus nucleocapsid protein, N, plays a central role in viral replication in encapsidating the three genomic RNA segments to form functional templates for transcription and replication by the viral RNA-dependent RNA polymerase. Here we report functional mapping of interacting domains of the Bunyamwera orthobunyavirus N protein by yeast and mammalian two-hybrid systems, immunoprecipitation experiments, and chemical cross-linking studies. N forms a range of multimers from dimers to high-molecular-weight structures, independently of the presence of RNA. Deletion of the N- or C-terminal domains resulted in loss of activity in a minireplicon assay and a decreased capacity for N to form higher multimers. Our data suggest a head-to-head and tail-to-tail multimerization model for the orthobunyavirus N protein.


* Corresponding author. Mailing address: Institute of Virology, Church Street, Glasgow G11 5JR, Scotland, United Kingdom. Phone: 44-141-330-4024. Fax: 44-141-337-2236. E-mail: r.elliott{at}vir.gla.ac.uk.

{dagger} Present address: Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, MN 55905.

{ddagger} Present address: Centre for Emergency Preparedness and Response, Health Protection Agency, Porton Down, Salisbury SP4 0JG, United Kingdom.


Journal of Virology, October 2005, p. 13166-13172, Vol. 79, No. 20
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.20.13166-13172.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.




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