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 Hardy, R. W.
Right arrow Articles by Wertz, G. W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hardy, R. W.
Right arrow Articles by Wertz, G. W.

 Previous Article  |  Next Article 

J. Virol., 01 1998, 520-526, Vol 72, No. 1
Copyright © 1998, American Society for Microbiology

The product of the respiratory syncytial virus M2 gene ORF1 enhances readthrough of intergenic junctions during viral transcription

RW Hardy and GW Wertz
Department of Microbiology, University of Alabama School of Medicine at Birmingham, 35294, USA.

The mRNA encoding the M2 protein of respiratory syncytial (RS) virus contains two open reading frames (ORFs). ORF1 encodes the 22-kDa structural protein, M2, and ORF2 has the potential to encode a 10-kDa protein (90 amino acids). Using a vaccinia virus T7 expression system, we examined the RNA synthetic activities of mono- and dicistronic subgenomic replicons of RS virus by direct metabolic labeling of RNA in the presence and absence of the products of ORF1 and ORF2. In the absence of ORF1 and ORF2, the negative- and positive-sense products of genomic RNA replication and positive-sense polyadenylated mRNA(s) were synthesized. Expression of the whole M2 transcription unit (containing ORF1 and ORF2) or ORF1 alone caused an increase in the synthesis of polyadenylated mRNA, the majority of which was due to a substantial increase in the quantity of polycistronic mRNAs generated by the polymerase failing to terminate at gene end signals. In agreement with previous reports, the ORF2 product was found to inhibit viral RNA replication and mRNA transcription. These data show that the M2 protein functions as a transcriptional antiterminator that enhances the ability of the viral RNA polymerase to read through intergenic junctions. The role of such a function during the viral life cycle is discussed.


This article has been cited by other articles:

  • Tran, T.-L., Castagne, N., Dubosclard, V., Noinville, S., Koch, E., Moudjou, M., Henry, C., Bernard, J., Yeo, R. P., Eleouet, J.-F. (2009). The Respiratory Syncytial Virus M2-1 Protein Forms Tetramers and Interacts with RNA and P in a Competitive Manner. J. Virol. 83: 6363-6374 [Abstract] [Full Text]  
  • Song, W., Liu, G., Bosworth, C. A., Walker, J. R., Megaw, G. A., Lazrak, A., Abraham, E., Sullender, W. M., Matalon, S. (2009). Respiratory Syncytial Virus Inhibits Lung Epithelial Na+ Channels by Up-regulating Inducible Nitric-oxide Synthase. J. Biol. Chem. 284: 7294-7306 [Abstract] [Full Text]  
  • de Graaf, M., Osterhaus, A. D. M. E., Fouchier, R. A. M., Holmes, E. C. (2008). Evolutionary dynamics of human and avian metapneumoviruses. J. Gen. Virol. 89: 2933-2942 [Abstract] [Full Text]  
  • Mohapatra, S. S., Boyapalle, S. (2008). Epidemiologic, Experimental, and Clinical Links between Respiratory Syncytial Virus Infection and Asthma. Clin. Microbiol. Rev. 21: 495-504 [Abstract] [Full Text]  
  • de Graaf, M., Herfst, S., Schrauwen, E. J. A., Choi, Y., van den Hoogen, B. G., Osterhaus, A. D. M. E., Fouchier, R. A. M. (2008). Specificity and functional interaction of the polymerase complex proteins of human and avian metapneumoviruses. J. Gen. Virol. 89: 975-983 [Abstract] [Full Text]  
  • Asenjo, A., Calvo, E., Villanueva, N. (2006). Phosphorylation of human respiratory syncytial virus P protein at threonine 108 controls its interaction with the M2-1 protein in the viral RNA polymerase complex. J. Gen. Virol. 87: 3637-3642 [Abstract] [Full Text]  
  • Hernandez-Sanchez, C., Bartulos, O., Valenciano, A. I., Mansilla, A., de Pablo, F. (2006). The regulated expression of chimeric tyrosine hydroxylase-insulin transcripts during early development. Nucleic Acids Res 34: 3455-3464 [Abstract] [Full Text]  
  • Cowton, V. M., McGivern, D. R., Fearns, R. (2006). Unravelling the complexities of respiratory syncytial virus RNA synthesis. J. Gen. Virol. 87: 1805-1821 [Abstract] [Full Text]  
  • Enterlein, S., Volchkov, V., Weik, M., Kolesnikova, L., Volchkova, V., Klenk, H.-D., Muhlberger, E. (2006). Rescue of Recombinant Marburg Virus from cDNA Is Dependent on Nucleocapsid Protein VP30. J. Virol. 80: 1038-1043 [Abstract] [Full Text]  
  • Akiva, P., Toporik, A., Edelheit, S., Peretz, Y., Diber, A., Shemesh, R., Novik, A., Sorek, R. (2006). Transcription-mediated gene fusion in the human genome. Genome Res 16: 30-36 [Abstract] [Full Text]  
  • Weik, M., Enterlein, S., Schlenz, K., Muhlberger, E. (2005). The Ebola Virus Genomic Replication Promoter Is Bipartite and Follows the Rule of Six. J. Virol. 79: 10660-10671 [Abstract] [Full Text]  
  • Buchholz, U. J., Biacchesi, S., Pham, Q. N., Tran, K. C., Yang, L., Luongo, C. L., Skiadopoulos, M. H., Murphy, B. R., Collins, P. L. (2005). Deletion of M2 Gene Open Reading Frames 1 and 2 of Human Metapneumovirus: Effects on RNA Synthesis, Attenuation, and Immunogenicity. J. Virol. 79: 6588-6597 [Abstract] [Full Text]  
  • Garcia-Barreno, B., Steel, J., Paya, M., Martinez-Sobrido, L., Delgado, T., Yeo, R. P., Melero, J. A. (2005). Epitope mapping of human respiratory syncytial virus 22K transcription antitermination factor: role of N-terminal sequences in protein folding. J. Gen. Virol. 86: 1103-1107 [Abstract] [Full Text]  
  • Asenjo, A., Rodriguez, L., Villanueva, N. (2005). Determination of phosphorylated residues from human respiratory syncytial virus P protein that are dynamically dephosphorylated by cellular phosphatases: a possible role for serine 54. J. Gen. Virol. 86: 1109-1120 [Abstract] [Full Text]  
  • Mason, S. W., Lawetz, C., Gaudette, Y., Do, F., Scouten, E., Lagace, L., Simoneau, B., Liuzzi, M. (2004). Polyadenylation-dependent screening assay for respiratory syncytial virus RNA transcriptase activity and identification of an inhibitor. Nucleic Acids Res 32: 4758-4767 [Abstract] [Full Text]  
  • Herfst, S., de Graaf, M., Schickli, J. H., Tang, R. S., Kaur, J., Yang, C.-F., Spaete, R. R., Haller, A. A., van den Hoogen, B. G., Osterhaus, A. D. M. E., Fouchier, R. A. M. (2004). Recovery of Human Metapneumovirus Genetic Lineages A and B from Cloned cDNA. J. Virol. 78: 8264-8270 [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]  
  • 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]  
  • Mason, S. W., Aberg, E., Lawetz, C., DeLong, R., Whitehead, P., Liuzzi, M. (2003). Interaction between Human Respiratory Syncytial Virus (RSV) M2-1 and P Proteins Is Required for Reconstitution of M2-1-Dependent RSV Minigenome Activity. J. Virol. 77: 10670-10676 [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]  
  • Zhou, H., Cheng, X., Jin, H. (2003). Identification of Amino Acids That Are Critical to the Processivity Function of Respiratory Syncytial Virus M2-1 Protein. J. Virol. 77: 5046-5053 [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]  
  • Krempl, C., Murphy, B. R., Collins, P. L. (2002). Recombinant Respiratory Syncytial Virus with the G and F Genes Shifted to the Promoter-Proximal Positions. J. Virol. 76: 11931-11942 [Abstract] [Full Text]  
  • Modrof, J., Muhlberger, E., Klenk, H.-D., Becker, S. (2002). Phosphorylation of VP30 Impairs Ebola Virus Transcription. J. Biol. Chem. 277: 33099-33104 [Abstract] [Full Text]  
  • Weik, M., Modrof, J., Klenk, H.-D., Becker, S., Muhlberger, E. (2002). Ebola Virus VP30-Mediated Transcription Is Regulated by RNA Secondary Structure Formation. J. Virol. 76: 8532-8539 [Abstract] [Full Text]  
  • Lu, B., Brazas, R., Ma, C.-H., Kristoff, T., Cheng, X., Jin, H. (2002). Identification of Temperature-Sensitive Mutations in the Phosphoprotein of Respiratory Syncytial Virus That Are Likely Involved in Its Interaction with the Nucleoprotein. J. Virol. 76: 2871-2880 [Abstract] [Full Text]  
  • Fearns, R., Peeples, M. E., Collins, P. L. (2002). Mapping the Transcription and Replication Promoters of Respiratory Syncytial Virus. J. Virol. 76: 1663-1672 [Abstract] [Full Text]  
  • Cartee, T. L., Wertz, G. W. (2001). Respiratory Syncytial Virus M2-1 Protein Requires Phosphorylation for Efficient Function and Binds Viral RNA during Infection. J. Virol. 75: 12188-12197 [Abstract] [Full Text]  
  • Gubbay, O., Curran, J., Kolakofsky, D. (2001). Sendai virus genome synthesis and assembly are coupled: a possible mechanism to promote viral RNA polymerase processivity. J. Gen. Virol. 82: 2895-2903 [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]  
  • Stope, M. B., Karger, A., Schmidt, U., Buchholz, U. J. (2001). Chimeric Bovine Respiratory Syncytial Virus with Attachment and Fusion Glycoproteins Replaced by Bovine Parainfluenza Virus Type 3 Hemagglutinin-Neuraminidase and Fusion Proteins. J. Virol. 75: 9367-9377 [Abstract] [Full Text]  
  • Bullido, R., Gómez-Puertas, P., Saiz, M. J., Portela, A. (2001). Influenza A Virus NEP (NS2 Protein) Downregulates RNA Synthesis of Model Template RNAs. J. Virol. 75: 4912-4917 [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]  
  • Harmon, S. B., Megaw, A. G., Wertz, G. W. (2001). RNA Sequences Involved in Transcriptional Termination of Respiratory Syncytial Virus. J. Virol. 75: 36-44 [Abstract] [Full Text]  
  • Cuesta, I., Geng, X., Asenjo, A., Villanueva, N. (2000). Structural Phosphoprotein M2-1 of the Human Respiratory Syncytial Virus Is an RNA Binding Protein. J. Virol. 74: 9858-9867 [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]  
  • Hardy, R. W., Wertz, G. W. (2000). The Cys3-His1 Motif of the Respiratory Syncytial Virus M2-1 Protein Is Essential for Protein Function. J. Virol. 74: 5880-5885 [Abstract] [Full Text]  
  • Fearns, R., Collins, P. L., Peeples, M. E. (2000). Functional Analysis of the Genomic and Antigenomic Promoters of Human Respiratory Syncytial Virus. J. Virol. 74: 6006-6014 [Abstract] [Full Text]  
  • Buchholz, U. J., Granzow, H., Schuldt, K., Whitehead, S. S., Murphy, B. R., Collins, P. L. (2000). Chimeric Bovine Respiratory Syncytial Virus with Glycoprotein Gene Substitutions from Human Respiratory Syncytial Virus (HRSV): Effects on Host Range and Evaluation as a Live-Attenuated HRSV Vaccine. J. Virol. 74: 1187-1199 [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]  
  • Peeples, M. E., Collins, P. L. (2000). Mutations in the 5' Trailer Region of a Respiratory Syncytial Virus Minigenome Which Limit RNA Replication to One Step. J. Virol. 74: 146-155 [Abstract] [Full Text]  
  • Villanueva, N., Hardy, R., Asenjo, A., Yu, Q., Wertz, G. (2000). The bulk of the phosphorylation of human respiratory syncytial virus phosphoprotein is not essential but modulates viral RNA transcription and replication. J. Gen. Virol. 81: 129-133 [Abstract] [Full Text]  
  • Dupuy, L. C., Dobson, S., Bitko, V., Barik, S. (1999). Casein Kinase 2-Mediated Phosphorylation of Respiratory Syncytial Virus Phosphoprotein P Is Essential for the Transcription Elongation Activity of the Viral Polymerase; Phosphorylation by Casein Kinase 1 Occurs Mainly at Ser215 and Is without Effect. J. Virol. 73: 8384-8392 [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]  
  • Ahmadian, G., Chambers, P., Easton, A. J. (1999). Detection and characterization of proteins encoded by the second ORF of the M2 gene of pneumoviruses. J. Gen. Virol. 80: 2011-2016 [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]  
  • Marriott, A. C., Wilson, S. D., Randhawa, J. S., Easton, A. J. (1999). A Single Amino Acid Substitution in the Phosphoprotein of Respiratory Syncytial Virus Confers Thermosensitivity in a Reconstituted RNA Polymerase System. J. Virol. 73: 5162-5165 [Abstract] [Full Text]  
  • Juhasz, K., Murphy, B. R., Collins, P. L. (1999). The Major Attenuating Mutations of the Respiratory Syncytial Virus Vaccine Candidate cpts530/1009 Specify Temperature-Sensitive Defects in Transcription and Replication and a Non-Temperature-Sensitive Alteration in mRNA Termination. J. Virol. 73: 5176-5180 [Abstract] [Full Text]  
  • Domachowske, J. B., Rosenberg, H. F. (1999). Respiratory Syncytial Virus Infection: Immune Response, Immunopathogenesis, and Treatment. Clin. Microbiol. Rev. 12: 298-309 [Abstract] [Full Text]  
  • Hardy, R. W., Harmon, S. B., Wertz, G. W. (1999). Diverse Gene Junctions of Respiratory Syncytial Virus Modulate the Efficiency of Transcription Termination and Respond Differently to M2-Mediated Antitermination. J. Virol. 73: 170-176 [Abstract] [Full Text]  
  • Buchholz, U. J., Finke, S., Conzelmann, K.-K. (1999). Generation of Bovine Respiratory Syncytial Virus (BRSV) from cDNA: BRSV NS2 Is Not Essential for Virus Replication in Tissue Culture, and the Human RSV Leader Region Acts as a Functional BRSV Genome Promoter. J. Virol. 73: 251-259 [Abstract] [Full Text]  
  • Teng, M. N., Collins, P. L. (1999). Altered Growth Characteristics of Recombinant Respiratory Syncytial Viruses Which Do Not Produce NS2 Protein. J. Virol. 73: 466-473 [Abstract] [Full Text]  
  • Muhlberger, E., Lotfering, B., Klenk, H.-D., Becker, S. (1998). Three of the Four Nucleocapsid Proteins of Marburg Virus, NP, VP35, and L, Are Sufficient To Mediate Replication and Transcription of Marburg Virus-Specific Monocistronic Minigenomes. J. Virol. 72: 8756-8764 [Abstract] [Full Text]  
  • Teng, M. N., Collins, P. L. (1998). Identification of the Respiratory Syncytial Virus Proteins Required for Formation and Passage of Helper-Dependent Infectious Particles. J. Virol. 72: 5707-5716 [Abstract] [Full Text]  
  • Magrangeas, F., Pitiot, G., Dubois, S., Bragado-Nilsson, E., Cherel, M., Jobert, S., Lebeau, B., Boisteau, O., Lethe, B., Mallet, J., Jacques, Y., Minvielle, S. (1998). Cotranscription and Intergenic Splicing of Human Galactose-1-phosphate Uridylyltransferase and Interleukin-11 Receptor alpha -Chain Genes Generate a Fusion mRNA in Normal Cells. IMPLICATION FOR THE PRODUCTION OF MULTIDOMAIN PROTEINS DURING EVOLUTION. J. Biol. Chem. 273: 16005-16010 [Abstract] [Full Text]