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Journal of Virology, August 2004, p. 8059-8067, Vol. 78, No. 15
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.15.8059-8067.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Interactions between the Inner and Outer Capsids of Bluetongue Virus

Emma L. Nason,1 Rosalba Rothagel,1 Sharmila K. Mukherjee,1 Alak Kanti Kar,2 Mario Forzan,3 B. V. Venkataram Prasad,4 and Polly Roy*

Verna and Marrs McLean Department of Biochemistry and Molecular Biology,1 W. M. Keck Center for Computational Biology, Baylor College of Medicine, Houston, Texas 77030,4 Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama 35294,2 Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London WC1E 7HT, United Kingdom3

Received 15 December 2003/ Accepted 15 March 2004

Bluetongue virus is a large and structurally complex virus composed of three concentric capsid layers that surround 10 segments of a double-stranded RNA genome. X-ray crystallographic analysis of the particles without the outer capsid layer has provided atomic structural details of VP3 and VP7, which form the inner two layers. However, limited structural information is available on the other five proteins in the virion—two of which are important for receptor recognition, hemagglutination, and membrane interaction—are in the outer layer, and the others, important for endogenous transcriptase activity are internal. Here we report the electron cryomicroscopy (cryo-EM) reconstruction of the mature particle, which shows that the outer layer has a unique non-T = 13 icosahedral organization consisting of two distinct triskelion and globular motifs interacting extensively with the underlying T = 13 layer. Comparative cryo-EM analysis of the recombinant corelike particles has shown that VP1 (viral polymerase) and VP4 (capping enzyme) together form a flower-shaped structure attached to the underside of VP3, directly beneath the fivefold axis. The structural data have been substantiated by biochemical studies demonstrating the interactions between the individual outer and inner capsid proteins.

* Corresponding author. Mailing address: Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel St., London WC1E 7HT, United Kingdom. Phone: 44 (0)20 7927 2324. Fax: 44 (0)207 7927 2839. E-mail: polly.roy{at}lshtm.ac.uk.

Journal of Virology, August 2004, p. 8059-8067, Vol. 78, No. 15
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.15.8059-8067.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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