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Journal of Virology, February 2000, p. 1994-2004, Vol. 74, No. 4
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Mutation of Predicted Virion Pit Residues Alters Binding of Theiler's Murine Encephalomyelitis Virus to BHK-21 Cells

Shannon Hertzler,1 Ming Luo,2 and Howard L. Lipton3,4,*

Integrated Graduate Program, Northwestern University Medical School,1 and Departments of Neurology, Microbiology-Immunology, and Biochemistry, Molecular Biology, and Cell Biology, Northwestern University,3 Chicago, and Division of Neurology, Evanston Hospital, Evanston,4 Illinois, and Center for Macromolecular Crystallography, University of Alabama, Birmingham, Alabama2

Received 4 August 1999/Accepted 22 November 1999

Theiler's murine encephalomyelitis virus (TMEV), a natural pathogen of mice, is a member of the genus Cardiovirus in the family Picornaviridae. Structural studies indicate that the cardiovirus pit, a deep depression on the surface of the virion, is involved in receptor attachment; however, this notion has never been systematically tested. Therefore, we used BeAn virus, a less virulent TMEV, to study the effect of site-specific mutation of selected pit amino acids on viral binding as well as other replicative functions of the virus. Four amino acids within the pit, V1091, P1153, A1225 and P3179, were selected for mutagenesis to evaluate their role in receptor attachment. Three amino acid replacements were made at each site, the first a conservative replacement, followed by progressively more radical amino acid changes in order to detect variable effects at each site. A total of seven viable mutant viruses were recovered and characterized for their binding properties to BHK-21 cells, capsid stability at 40°C, viral RNA replication, single- and multistep growth kinetics, and virus translation. Our data implicate three of these residues in TMEV-cell receptor attachment.

* Corresponding author. Mailing address: Division of Neurology, Evanston Hospital, 2650 Ridge Ave., Evanston, IL 60201-1782. Phone: (847) 570-2168. Fax: (847) 570-1568. E-mail: hllipton{at}merle.acns.nwu.edu.

Journal of Virology, February 2000, p. 1994-2004, Vol. 74, No. 4
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Reddi, H. V., Kallio, P., Lipton, H. L. (2003). Galactose is needed only for expression of co-receptors used by Theiler's murine encephalomyelitis virus as the virus does not directly bind galactose or use the UDP-galactose transporter as a receptor. J. Gen. Virol. 84: 845-849 [Abstract] [Full Text]  
  • Kumar, A. S. M., Kallio, P., Luo, M., Lipton, H. L. (2003). Amino Acid Substitutions in VP2 Residues Contacting Sialic Acid in Low-Neurovirulence BeAn Virus Dramatically Reduce Viral Binding and Spread of Infection. J. Virol. 77: 2709-2716 [Abstract] [Full Text]  
  • Jnaoui, K., Minet, M., Michiels, T. (2002). Mutations That Affect the Tropism of DA and GDVII Strains of Theiler's Virus In Vitro Influence Sialic Acid Binding and Pathogenicity. J. Virol. 76: 8138-8147 [Abstract] [Full Text]  
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