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 Oomens, A. G. P.
Right arrow Articles by Wertz, G. W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Oomens, A. G. P.
Right arrow Articles by Wertz, G. W.

 Previous Article  |  Next Article 

Journal of Virology, March 2003, p. 3785-3798, Vol. 77, No. 6
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.6.3785-3798.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Infectivity of a Human Respiratory Syncytial Virus Lacking the SH, G, and F Proteins Is Efficiently Mediated by the Vesicular Stomatitis Virus G Protein

A. G. P. Oomens, A. G. Megaw, and G. W. Wertz*

University of Alabama School of Medicine, Birmingham, Alabama 35294

Received 4 October 2002/ Accepted 11 November 2002

To examine the requirements of the human respiratory syncytial virus (HRSV) SH (small hydrophobic), G (attachment), and F (fusion) proteins for virus infectivity and morphology, we used the prototype A2 strain of HRSV to generate a series of cDNAs from which (i) the SH open reading frame (ORF), (ii) the SH and G ORFs, or (iii) the SH, G, and F ORFs were deleted. Each deleted ORF was replaced as follows: the SH ORF was replaced with that of green fluorescent protein; the G ORF was replaced with that of Gvsv, a chimeric glycoprotein consisting of the vesicular stomatitis Indiana virus (VSIV) G protein ecto- and transmembrane domains coupled to the HRSV F cytoplasmic tail; and the F ORF was replaced with that of marker protein ß-glucuronidase. The number of genes and the intergenic junctions in the constructs were kept as found in A2 virus in order to maintain authentic levels of transcription. Infectious viruses were recovered from all three engineered cDNAs and designated RS{Delta}sh, RS{Delta}sh,g/Gvsv, and RS{Delta}sh,g,f/Gvsv, respectively. Low-pH-induced syncytium formation was observed in cells infected with viruses RS{Delta}SH,G/Gvsv and RS{Delta}SH,G,F/Gvsv, indicating that Gvsv was expressed and functional. Neutralization of infectivity by anti-VSIV G antibodies and inhibition of entry by ammonium chloride showed that RS{Delta}SH,G,F/Gvsv infectivity was mediated by Gvsv and that an acidification step was required for entry into the host cell, similar to VSIV virions. All three engineered viruses displayed growth kinetics and virus yields similar to a wild-type A2 virus, both in Vero and HEp-2 cells. Abundant virus-induced filaments were observed at the surface of cells infected with each of the three engineered viruses or with virus A2, indicating that neither the SH and G proteins nor the F protein ecto- and transmembrane domains were required for the formation of these structures. This is the first report of the recovery of an infectious HRSV lacking a fusion protein of the Paramyxoviridae family and of manipulation of the HRSV entry pathway via incorporation of a nonparamyxoviral transmembrane glycoprotein.

* Corresponding author. Mailing address: Department of Microbiology, The Medical School, University of Alabama at Birmingham, BBRB 17 Room 366, 845 19th St. S., Birmingham, AL 35294. Phone: (205) 934-0877. Fax: (205) 934-1636. E-mail: gailw{at}uab.edu.

Journal of Virology, March 2003, p. 3785-3798, Vol. 77, No. 6
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.6.3785-3798.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Basu, D., Walkiewicz, M. P., Frieman, M., Baric, R. S., Auble, D. T., Engel, D. A. (2009). Novel Influenza Virus NS1 Antagonists Block Replication and Restore Innate Immune Function. J. Virol. 83: 1881-1891 [Abstract] [Full Text]  
  • Batonick, M., Oomens, A. G. P., Wertz, G. W. (2008). Human Respiratory Syncytial Virus Glycoproteins Are Not Required for Apical Targeting and Release from Polarized Epithelial Cells. J. Virol. 82: 8664-8672 [Abstract] [Full Text]  
  • Fleming, E. H., Kolokoltsov, A. A., Davey, R. A., Nichols, J. E., Roberts, N. J. Jr. (2006). Respiratory Syncytial Virus F Envelope Protein Associates with Lipid Rafts without a Requirement for Other Virus Proteins. J. Virol. 80: 12160-12170 [Abstract] [Full Text]  
  • Oomens, A. G. P., Bevis, K. P., Wertz, G. W. (2006). The Cytoplasmic Tail of the Human Respiratory Syncytial Virus F Protein Plays Critical Roles in Cellular Localization of the F Protein and Infectious Progeny Production. J. Virol. 80: 10465-10477 [Abstract] [Full Text]  
  • Gower, T. L., Pastey, M. K., Peeples, M. E., Collins, P. L., McCurdy, L. H., Hart, T. K., Guth, A., Johnson, T. R., Graham, B. S. (2005). RhoA Signaling Is Required for Respiratory Syncytial Virus-Induced Syncytium Formation and Filamentous Virion Morphology. J. Virol. 79: 5326-5336 [Abstract] [Full Text]  
  • Martinez, I., Wertz, G. W. (2005). Biological Differences between Vesicular Stomatitis Virus Indiana and New Jersey Serotype Glycoproteins: Identification of Amino Acid Residues Modulating pH-Dependent Infectivity. J. Virol. 79: 3578-3585 [Abstract] [Full Text]  
  • Oomens, A. G. P., Wertz, G. W. (2004). trans-Complementation Allows Recovery of Human Respiratory Syncytial Viruses That Are Infectious but Deficient in Cell-to-Cell Transmission. J. Virol. 78: 9064-9072 [Abstract] [Full Text]  
  • Johnson, K. L., Price, B. D., Eckerle, L. D., Ball, L. A. (2004). Nodamura Virus Nonstructural Protein B2 Can Enhance Viral RNA Accumulation in both Mammalian and Insect Cells. J. Virol. 78: 6698-6704 [Abstract] [Full Text]  
  • Rodriguez, L., Cuesta, I., Asenjo, A., Villanueva, N. (2004). Human respiratory syncytial virus matrix protein is an RNA-binding protein: binding properties, location and identity of the RNA contact residues. J. Gen. Virol. 85: 709-719 [Abstract] [Full Text]  
  • Oomens, A. G. P., Wertz, G. W. (2004). The Baculovirus GP64 Protein Mediates Highly Stable Infectivity of a Human Respiratory Syncytial Virus Lacking Its Homologous Transmembrane Glycoproteins. J. Virol. 78: 124-135 [Abstract] [Full Text]  
  • Cartee, T. L., Megaw, A. G., Oomens, A. G. P., Wertz, G. W. (2003). Identification of a Single Amino Acid Change in the Human Respiratory Syncytial Virus L Protein That Affects Transcriptional Termination. J. Virol. 77: 7352-7360 [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»