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 Graff, J.
Right arrow Articles by Emerson, S. U.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Graff, J.
Right arrow Articles by Emerson, S. U.

 Previous Article  |  Next Article 

Journal of Virology, February 2008, p. 1185-1194, Vol. 82, No. 3
0022-538X/08/$08.00+0     doi:10.1128/JVI.01219-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Mutations within Potential Glycosylation Sites in the Capsid Protein of Hepatitis E Virus Prevent the Formation of Infectious Virus Particles{triangledown}

Judith Graff,1 Yi-Hua Zhou,1 Udana Torian,1 Hanh Nguyen,1 Marisa St. Claire,3 Claro Yu,2 Robert H. Purcell,2 and Suzanne U. Emerson1*

Molecular Hepatitis,1 Hepatitis Viruses Sections, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892,2 Bioqual, Incorporated, 2501 Research Boulevard, Rockville, Maryland 208503

Received 4 June 2007/ Accepted 11 November 2007

Hepatitis E virus is a nonenveloped RNA virus. However, the single capsid protein resembles a typical glycoprotein in that it contains a signal sequence and potential glycosylation sites that are utilized when recombinant capsid protein is overexpressed in cell culture. In order to determine whether these unexpected observations were biologically relevant or were artifacts of overexpression, we analyzed capsid protein produced during a normal viral replication cycle. In vitro transcripts from an infectious cDNA clone mutated to eliminate potential glycosylation sites were transfected into cultured Huh-7 cells and into the livers of rhesus macaques. The mutations did not detectably affect genome replication or capsid protein synthesis in cell culture. However, none of the mutants infected rhesus macaques. Velocity sedimentation analyses of transfected cell lysates revealed that mutation of the first two glycosylation sites prevented virion assembly, whereas mutation of the third site permitted particle formation and RNA encapsidation, but the particles were not infectious. However, conservative mutations that did not destroy glycosylation motifs also prevented infection. Overall, the data suggested that the mutations were lethal because they perturbed protein structure rather than because they eliminated glycosylation.

* Corresponding author. Mailing address: Molecular Hepatitis Section, LID, NIAID, National Institutes of Health, Room 6537, Building 50, 50 South Drive, MSC 8009, Bethesda, MD 20892-8009. Phone: (301) 496-2787. Fax: (301) 402-0524. E-mail: semerson{at}niaid.nih.gov

{triangledown} Published ahead of print on 21 November 2007.

Journal of Virology, February 2008, p. 1185-1194, Vol. 82, No. 3
0022-538X/08/$08.00+0     doi:10.1128/JVI.01219-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Kalia, M., Chandra, V., Rahman, S. A., Sehgal, D., Jameel, S. (2009). Heparan Sulfate Proteoglycans Are Required for Cellular Binding of the Hepatitis E Virus ORF2 Capsid Protein and for Viral Infection. J. Virol. 83: 12714-12724 [Abstract] [Full Text]  
  • Yamashita, T., Mori, Y., Miyazaki, N., Cheng, R. H., Yoshimura, M., Unno, H., Shima, R., Moriishi, K., Tsukihara, T., Li, T. C., Takeda, N., Miyamura, T., Matsuura, Y. (2009). Biological and immunological characteristics of hepatitis E virus-like particles based on the crystal structure. Proc. Natl. Acad. Sci. USA 106: 12986-12991 [Abstract] [Full Text]  
  • Bilic, I., Jaskulska, B., Basic, A., Morrow, C. J., Hess, M. (2009). Sequence analysis and comparison of avian hepatitis E viruses from Australia and Europe indicate the existence of different genotypes. J. Gen. Virol. 90: 863-873 [Abstract] [Full Text]  
  • Yamada, K., Takahashi, M., Hoshino, Y., Takahashi, H., Ichiyama, K., Tanaka, T., Okamoto, H. (2009). Construction of an infectious cDNA clone of hepatitis E virus strain JE03-1760F that can propagate efficiently in cultured cells. J. Gen. Virol. 90: 457-462 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»