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 Teng, M. N.
Right arrow Articles by Collins, P. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Teng, M. N.
Right arrow Articles by Collins, P. L.

 Previous Article  |  Next Article 

Journal of Virology, January 1999, p. 466-473, Vol. 73, No. 1
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Altered Growth Characteristics of Recombinant Respiratory Syncytial Viruses Which Do Not Produce NS2 Protein

Michael N. Teng and Peter L. Collins*

Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892-0720

Received 10 July 1998/Accepted 16 September 1998

The second gene in the 3'-to-5' gene order in respiratory syncytial virus (RSV) encodes the nonstructural protein NS2, for which there is no assigned function. To study the function of NS2, we have used a recently developed reverse genetics system to ablate expression of NS2 in recombinant RSV. A full-length cDNA copy of the antigenome of RSV A2 strain under the control of a T7 promoter was modified by introduction of tandem termination codons within the NS2 open reading frame (NS2stop) or by deletion of the entire NS2 gene (Delta NS2). The NS2 knockout antigenomic cDNAs were cotransfected with plasmids encoding the N, P, L, and M2-1 proteins of RSV, each controlled by the T7 promoter, into cells infected with a vaccinia virus recombinant expressing T7 RNA polymerase. Recombinant NS2stop and Delta NS2 RSVs were recovered and characterized. Both types of NS2 knockout virus displayed pinpoint plaque morphology and grew more slowly than wild-type RSV. The expression of monocistronic mRNAs for the five genes examined (NS1, NS2, N, F, and L) was unchanged in cells infected with either type of NS2 knockout virus, except that no NS2 mRNA was detected with the Delta NS2 virus. Synthesis of readthrough mRNAs was affected only for the Delta NS2 virus, where the NS1-NS2, NS2-N, and NS1-NS2-N mRNAs were replaced with the predicted novel NS1-N mRNA. Upon passage, the NS2stop virus stock rapidly developed revertants which expressed NS2 protein and grew with similar plaque morphology and kinetics wild-type RSV. Sequence analysis confirmed that the termination codons had reverted to sense, albeit not the wild-type assignments, and provided evidence consistent with biased hypermutation. No revertants were recovered from recombinant Delta NS2 RSV. These results show that the NS2 protein is not essential for RSV replication, although its presence greatly improves virus growth in cell culture. The attenuated phenotype of these mutant viruses, coupled with the expected genetic stability associated with gene deletions, suggests that the Delta NS2 RSV is a candidate for vaccine development.

* Corresponding author. Mailing address: Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, 7 Center Dr. MSC 0720, Bethesda, MD 20892-0720. Phone: (301) 496-4205. Fax: (301) 496-8312. E-mail: pcollins{at}atlas.niaid.nih.gov.

Journal of Virology, January 1999, p. 466-473, Vol. 73, No. 1
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Swedan, S., Musiyenko, A., Barik, S. (2009). Respiratory Syncytial Virus Nonstructural Proteins Decrease Levels of Multiple Members of the Cellular Interferon Pathways. J. Virol. 83: 9682-9693 [Abstract] [Full Text]  
  • Ghildyal, R., Ho, A., Dias, M., Soegiyono, L., Bardin, P. G., Tran, K. C., Teng, M. N., Jans, D. A. (2009). The Respiratory Syncytial Virus Matrix Protein Possesses a Crm1-Mediated Nuclear Export Mechanism. J. Virol. 83: 5353-5362 [Abstract] [Full Text]  
  • Ling, Z., Tran, K. C., Teng, M. N. (2009). Human Respiratory Syncytial Virus Nonstructural Protein NS2 Antagonizes the Activation of Beta Interferon Transcription by Interacting with RIG-I. J. Virol. 83: 3734-3742 [Abstract] [Full Text]  
  • Buchholz, U. J., Ward, J. M., Lamirande, E. W., Heinze, B., Krempl, C. D., Collins, P. L. (2009). Deletion of Nonstructural Proteins NS1 and NS2 from Pneumonia Virus of Mice Attenuates Viral Replication and Reduces Pulmonary Cytokine Expression and Disease. J. Virol. 83: 1969-1980 [Abstract] [Full Text]  
  • Munir, S., Le Nouen, C., Luongo, C., Buchholz, U. J., Collins, P. L., Bukreyev, A. (2008). Nonstructural Proteins 1 and 2 of Respiratory Syncytial Virus Suppress Maturation of Human Dendritic Cells. J. Virol. 82: 8780-8796 [Abstract] [Full Text]  
  • McCann, K. L., Imani, F. (2007). Transforming Growth Factor {beta} Enhances Respiratory Syncytial Virus Replication and Tumor Necrosis Factor Alpha Induction in Human Epithelial Cells. J. Virol. 81: 2880-2886 [Abstract] [Full Text]  
  • Bitko, V., Shulyayeva, O., Mazumder, B., Musiyenko, A., Ramaswamy, M., Look, D. C., Barik, S. (2007). Nonstructural Proteins of Respiratory Syncytial Virus Suppress Premature Apoptosis by an NF-{kappa}B-Dependent, Interferon-Independent Mechanism and Facilitate Virus Growth. J. Virol. 81: 1786-1795 [Abstract] [Full Text]  
  • Bukreyev, A., Skiadopoulos, M. H., Murphy, B. R., Collins, P. L. (2006). Nonsegmented negative-strand viruses as vaccine vectors.. J. Virol. 80: 10293-10306 [Full Text]  
  • Kotelkin, A., Belyakov, I. M., Yang, L., Berzofsky, J. A., Collins, P. L., Bukreyev, A. (2006). The NS2 Protein of Human Respiratory Syncytial Virus Suppresses the Cytotoxic T-Cell Response as a Consequence of Suppressing the Type I Interferon Response.. J. Virol. 80: 5958-5967 [Abstract] [Full Text]  
  • Cheng, X., Park, H., Zhou, H., Jin, H. (2005). Overexpression of the M2-2 Protein of Respiratory Syncytial Virus Inhibits Viral Replication. J. Virol. 79: 13943-13952 [Abstract] [Full Text]  
  • Spann, K. M., Tran, K. C., Collins, P. L. (2005). Effects of Nonstructural Proteins NS1 and NS2 of Human Respiratory Syncytial Virus on Interferon Regulatory Factor 3, NF-{kappa}B, and Proinflammatory Cytokines. J. Virol. 79: 5353-5362 [Abstract] [Full Text]  
  • Thoulouze, M.-I., Bouguyon, E., Carpentier, C., Bremont, M. (2004). Essential Role of the NV Protein of Novirhabdovirus for Pathogenicity in Rainbow Trout. J. Virol. 78: 4098-4107 [Abstract] [Full Text]  
  • Spann, K. M., Tran, K.-C., Chi, B., Rabin, R. L., Collins, P. L. (2004). Suppression of the Induction of Alpha, Beta, and Gamma Interferons by the NS1 and NS2 Proteins of Human Respiratory Syncytial Virus in Human Epithelial Cells and Macrophages. J. Virol. 78: 4363-4369 [Abstract] [Full Text]  
  • Easton, A. J., Domachowske, J. B., Rosenberg, H. F. (2004). Animal Pneumoviruses: Molecular Genetics and Pathogenesis. Clin. Microbiol. Rev. 17: 390-412 [Abstract] [Full Text]  
  • Tran, K. C., Collins, P. L., Teng, M. N. (2004). Effects of Altering the Transcription Termination Signals of Respiratory Syncytial Virus on Viral Gene Expression and Growth In Vitro and In Vivo. J. Virol. 78: 692-699 [Abstract] [Full Text]  
  • Spann, K. M., Collins, P. L., Teng, M. N. (2003). Genetic Recombination during Coinfection of Two Mutants of Human Respiratory Syncytial Virus. J. Virol. 77: 11201-11211 [Abstract] [Full Text]  
  • Bossert, B., Marozin, S., Conzelmann, K.-K. (2003). Nonstructural Proteins NS1 and NS2 of Bovine Respiratory Syncytial Virus Block Activation of Interferon Regulatory Factor 3. J. Virol. 77: 8661-8668 [Abstract] [Full Text]  
  • Valarcher, J.-F., Furze, J., Wyld, S., Cook, R., Conzelmann, K.-K., Taylor, G. (2003). Role of Alpha/Beta Interferons in the Attenuation and Immunogenicity of Recombinant Bovine Respiratory Syncytial Viruses Lacking NS Proteins. J. Virol. 77: 8426-8439 [Abstract] [Full Text]  
  • Neumann, G., Whitt, M. A., Kawaoka, Y. (2002). A decade after the generation of a negative-sense RNA virus from cloned cDNA - what have we learned?. J. Gen. Virol. 83: 2635-2662 [Abstract] [Full Text]  
  • Teng, M. N., Collins, P. L. (2002). The Central Conserved Cystine Noose of the Attachment G Protein of Human Respiratory Syncytial Virus Is Not Required for Efficient Viral Infection In Vitro or In Vivo. J. Virol. 76: 6164-6171 [Abstract] [Full Text]  
  • Bossert, B., Conzelmann, K.-K. (2002). Respiratory Syncytial Virus (RSV) Nonstructural (NS) Proteins as Host Range Determinants: a Chimeric Bovine RSV with NS Genes from Human RSV Is Attenuated in Interferon-Competent Bovine Cells. J. Virol. 76: 4287-4293 [Abstract] [Full Text]  
  • Schmidt, A. C., Wenzke, D. R., McAuliffe, J. M., St. Claire, M., Elkins, W. R., Murphy, B. R., Collins, P. L. (2002). Mucosal Immunization of Rhesus Monkeys against Respiratory Syncytial Virus Subgroups A and B and Human Parainfluenza Virus Type 3 by Using a Live cDNA-Derived Vaccine Based on a Host Range-Attenuated Bovine Parainfluenza Virus Type 3 Vector Backbone. J. Virol. 76: 1089-1099 [Abstract] [Full Text]  
  • Tang, R. S., Nguyen, N., Cheng, X., Jin, H. (2001). Requirement of Cysteines and Length of the Human Respiratory Syncytial Virus M2-1 Protein for Protein Function and Virus Viability. J. Virol. 75: 11328-11335 [Abstract] [Full Text]  
  • Karger, A., Schmidt, U., Buchholz, U. J. (2001). Recombinant bovine respiratory syncytial virus with deletions of the G or SH genes: G and F proteins bind heparin. J. Gen. Virol. 82: 631-640 [Abstract] [Full Text]  
  • Bukreyev, A., Murphy, B. R., Collins, P. L. (2000). Respiratory Syncytial Virus Can Tolerate an Intergenic Sequence of at Least 160 Nucleotides with Little Effect on Transcription or Replication In Vitro and In Vivo. J. Virol. 74: 11017-11026 [Abstract] [Full Text]  
  • Teng, M. N., Whitehead, S. S., Bermingham, A., St. Claire, M., Elkins, W. R., Murphy, B. R., Collins, P. L. (2000). Recombinant Respiratory Syncytial Virus That Does Not Express the NS1 or M2-2 Protein Is Highly Attenuated and Immunogenic in Chimpanzees. J. Virol. 74: 9317-9321 [Abstract] [Full Text]  
  • Schlender, J., Bossert, B., Buchholz, U., Conzelmann, K.-K. (2000). Bovine Respiratory Syncytial Virus Nonstructural Proteins NS1 and NS2 Cooperatively Antagonize Alpha/Beta Interferon-Induced Antiviral Response. J. Virol. 74: 8234-8242 [Abstract] [Full Text]  
  • Tripp, R. A., Jones, L., Anderson, L. J., Brown, M. P. (2000). CD40 Ligand (CD154) Enhances the Th1 and Antibody Responses to Respiratory Syncytial Virus in the BALB/c Mouse. J. Immunol. 164: 5913-5921 [Abstract] [Full Text]  
  • Vialat, P., Billecocq, A., Kohl, A., Bouloy, M. (2000). The S Segment of Rift Valley Fever Phlebovirus (Bunyaviridae) Carries Determinants for Attenuation and Virulence in Mice. J. Virol. 74: 1538-1543 [Abstract] [Full Text]  
  • Jin, H., Cheng, X., Zhou, H. Z. Y., Li, S., Seddiqui, A. (2000). Respiratory Syncytial Virus That Lacks Open Reading Frame 2 of the M2 Gene (M2-2) Has Altered Growth Characteristics and Is Attenuated in Rodents. J. Virol. 74: 74-82 [Abstract] [Full Text]  
  • Bermingham, A., Collins, P. L. (1999). The M2-2 protein of human respiratory syncytial virus is a regulatory factor involved in the balance between RNA replication and transcription. Proc. Natl. Acad. Sci. USA 96: 11259-11264 [Abstract] [Full Text]  
  • Fearns, R., Collins, P. L. (1999). Role of the M2-1 Transcription Antitermination Protein of Respiratory Syncytial Virus in Sequential Transcription. J. Virol. 73: 5852-5864 [Abstract] [Full Text]  
  • Whitehead, S. S., Bukreyev, A., Teng, M. N., Firestone, C.-Y., St. Claire, M., Elkins, W. R., Collins, P. L., Murphy, B. R. (1999). Recombinant Respiratory Syncytial Virus Bearing a Deletion of either the NS2 or SH Gene Is Attenuated in Chimpanzees. J. Virol. 73: 3438-3442 [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»