JVI Figure table search 04
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Chandran, K.
Right arrow Articles by Nibert, M. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Chandran, K.
Right arrow Articles by Nibert, M. L.

 Previous Article  |  Next Article 

Journal of Virology, June 2001, p. 5335-5342, Vol. 75, No. 11
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.11.5335-5342.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Complete In Vitro Assembly of the Reovirus Outer Capsid Produces Highly Infectious Particles Suitable for Genetic Studies of the Receptor-Binding Protein

Kartik Chandran,1,dagger Xing Zhang,2 Norman H. Olson,2 Stephen B. Walker,2 James D. Chappell,3 Terence S. Dermody,3 Timothy S. Baker,2 and Max L. Nibert1,*

Department of Biochemistry and Institute for Molecular Virology, University of Wisconsin---Madison, Madison, Wisconsin 537061; Department of Biological Sciences, Purdue University, West Lafayette, Indiana 479072; and Departments of Pediatrics and Microbiology and Immunology and Elizabeth B. Lamb Center for Pediatric Research, Vanderbilt University School of Medicine, Nashville, Tennessee 372323

Received 15 November 2000/Accepted 7 March 2001

Mammalian reoviruses, prototype members of the Reoviridae family of nonenveloped double-stranded RNA viruses, use at least three proteins---sigma 1, µ1, and sigma 3---to enter host cells. sigma 1, a major determinant of cell tropism, mediates viral attachment to cellular receptors. Studies of sigma 1 functions in reovirus entry have been restricted by the lack of methodologies to produce infectious virions containing engineered mutations in viral proteins. To mitigate this problem, we produced virion-like particles by "recoating" genome-containing core particles that lacked sigma 1, µ1, and sigma 3 with recombinant forms of these proteins in vitro. Image reconstructions from cryoelectron micrographs of the recoated particles revealed that they closely resembled native virions in three-dimensional structure, including features attributable to sigma 1. The recoated particles bound to and infected cultured cells in a sigma 1-dependent manner and were approximately 1 million times as infectious as cores and 0.5 times as infectious as native virions. Experiments with recoated particles containing recombinant sigma 1 from either of two different reovirus strains confirmed that differences in cell attachment and infectivity previously observed between those strains are determined by the sigma 1 protein. Additional experiments showed that recoated particles containing sigma 1 proteins with engineered mutations can be used to analyze the effects of such mutations on the roles of particle-bound sigma 1 in infection. The results demonstrate a powerful new system for molecular genetic dissections of sigma 1 with respect to its structure, assembly into particles, and roles in entry.

* Corresponding author. Present address: Department of Microbiology and Molecular Genetics, Harvard Medical School, 200 Longwood Ave., Boston, MA 02115. Phone: (617) 432-4829. Fax: (617) 738-7664. E-mail: mnibert{at}hms.harvard.edu.

dagger Present address: Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Mass.

Journal of Virology, June 2001, p. 5335-5342, Vol. 75, No. 11
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.11.5335-5342.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:



Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
J. Bacteriol. Mol. Cell. Biol. Microbiol. Mol. Biol. Rev.
Clin. Vaccine Immunol. ALL ASM JOURNALS

Copyright © 2001 by the American Society for Microbiology. All rights reserved.