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 Fearns, R.
Right arrow Articles by Collins, P. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fearns, R.
Right arrow Articles by Collins, P. L.

 Previous Article  |  Next Article 

Journal of Virology, January 1999, p. 388-397, Vol. 73, No. 1
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Model for Polymerase Access to the Overlapped L Gene of Respiratory Syncytial Virus

Rachel Fearns and Peter L. Collins*

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

Received 21 April 1998/Accepted 23 September 1998

The last two genes of respiratory syncytial virus (RSV), M2 and L, overlap by 68 nucleotides, an arrangement which has counterparts in a number of nonsegmented negative-strand RNA viruses. Thus, the gene-end (GE) signal of M2 lies downstream of the L gene-start (GS) signal, separated by 45 nucleotides. Since RSV transcription ostensibly is sequential and unidirectional from a single promoter within the 3' leader region, it was unclear how the polymerase accesses the L GS signal. Furthermore, it was previously shown that 90% of transcripts which are initiated at the L GS signal are polyadenylated and terminated at the M2 GE signal, yielding a short, truncated L mRNA as the major transcription product of the L gene. Despite these apparent down-regulatory features, we show that the accumulation of full-length L mRNA during RSV infection is only sixfold less than that of its upstream neighbor, M2. We used cDNA-encoded genome analogs in an intracellular transcription assay to investigate the mechanism of transcription of the overlapped genes. Expression of L was found to be dependent on sequential transcription from the 3' end of the genome. Apart from the L GS signal, the only other strict requirement for initiation at L was the M2 GE signal. This implies that the polymerase accesses the L GS signal only following arrival at the M2 GE signal. Thus, polymerase which terminates at the M2 GE signal presumably scans upstream to initiate at the L GS signal. This also would provide a mechanism whereby polymerase which terminates prematurely during transcription of L could recycle from the M2 GE signal to the L GS signal, thereby accounting for the unexpectedly high level of synthesis of full-length L mRNA. The sequence and spacing between the two signals were not critical. Furthermore, the polymerase also was capable of efficiently transcribing from an L GS signal placed downstream of the M2 GE signal, implying that the overlapping arrangement is not obligatory. When copies of the L GS signal were placed concurrently upstream and downstream of the M2 GE signal, both were utilized. This finding indicates that a polymerase situated at a GE signal is capable of scanning for a GS signal in either the upstream or downstream direction and thereafter initiating transcription.

* Corresponding author. Mailing address: LID, NIAID, NIH, 7 Center Dr., MSC 0720, Bethesda, MD 20892-0720. Phone: (301) 496-3481. Fax: (301) 496-8312. E-mail: pcollins{at}atlas.niaid.nih.gov.

Journal of Virology, January 1999, p. 388-397, 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:

  • Poenisch, M., Wille, S., Staeheli, P., Schneider, U. (2008). Polymerase Read-Through at the First Transcription Termination Site Contributes to Regulation of Borna Disease Virus Gene Expression. J. Virol. 82: 9537-9545 [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]  
  • Hoffman, M. A., Thorson, L. M., Vickman, J. E., Anderson, J. S., May, N. A., Schweitzer, M. N. (2006). Roles of human parainfluenza virus type 3 bases 13 to 78 in replication and transcription: identification of an additional replication promoter element and evidence for internal transcription initiation.. J. Virol. 80: 5388-5396 [Abstract] [Full Text]  
  • Cordey, S., Roux, L. (2006). Transcribing paramyxovirus RNA polymerase engages the template at its 3' extremity.. J. Gen. Virol. 87: 665-672 [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]  
  • Cowton, V. M., Fearns, R. (2005). Evidence that the Respiratory Syncytial Virus Polymerase Is Recruited to Nucleotides 1 to 11 at the 3' End of the Nucleocapsid and Can Scan To Access Internal Signals. J. Virol. 79: 11311-11322 [Abstract] [Full Text]  
  • Agrawal, A., Tripp, R. A., Anderson, L. J., Nie, S. (2005). Real-Time Detection of Virus Particles and Viral Protein Expression with Two-Color Nanoparticle Probes. J. Virol. 79: 8625-8628 [Abstract] [Full Text]  
  • McGivern, D. R., Collins, P. L., Fearns, R. (2005). Identification of Internal Sequences in the 3' Leader Region of Human Respiratory Syncytial Virus That Enhance Transcription and Confer Replication Processivity. J. Virol. 79: 2449-2460 [Abstract] [Full Text]  
  • Vulliemoz, D., Cordey, S., Mottet-Osman, G., Roux, L. (2005). Nature of a paramyxovirus replication promoter influences a nearby transcription signal. J. Gen. Virol. 86: 171-180 [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]  
  • Le Mercier, P., Garcin, D., Garcia, E., Kolakofsky, D. (2003). Competition between the Sendai Virus N mRNA Start Site and the Genome 3'-End Promoter for Viral RNA Polymerase. J. Virol. 77: 9147-9155 [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]  
  • 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]  
  • Marriott, A. C., Smith, J. M., Easton, A. J. (2001). Fidelity of Leader and Trailer Sequence Usage by the Respiratory Syncytial Virus and Avian Pneumovirus Replication Complexes. J. Virol. 75: 6265-6272 [Abstract] [Full Text]  
  • Vulliémoz, D., Roux, L. (2001). "Rule of Six": How Does the Sendai Virus RNA Polymerase Keep Count?. J. Virol. 75: 4506-4518 [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]  
  • Whelan, S. P. J., Barr, J. N., Wertz, G. W. (2000). Identification of a Minimal Size Requirement for Termination of Vesicular Stomatitis Virus mRNA: Implications for the Mechanism of Transcription. J. Virol. 74: 8268-8276 [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]  


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