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Journal of Virology, September 2005, p. 11311-11322, Vol. 79, No. 17
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.17.11311-11322.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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

Vanessa M. Cowton and Rachel Fearns*

Division of Pathology and Neuroscience, University of Dundee, Dundee, DD1 9SY United Kingdom

Received 25 February 2005/ Accepted 9 June 2005

The 3'-terminal end of the respiratory syncytial virus genomic RNA contains a 44-nucleotide leader (Le) region adjoining the gene start signal of the first gene. Previous mapping studies demonstrated that there is a promoter located at the 3' end of Le, which can signal initiation of antigenome synthesis. The aim of this study was to investigate the role of the 3' terminus of the RNA template in (i) promoter recognition and (ii) determining the initiation site for antigenome synthesis. A panel of minigenomes containing additional sequence at the 3' end of the Le were analyzed for their ability to direct antigenome and mRNA synthesis. Minigenomes containing heterologous extensions of 6 nucleotides or more were unable to support efficient RNA synthesis. However, the activity of a minigenome with a 56-nucleotide extension could be restored by insertion of Le nucleotides 1 to 11 or 1 to 13 at the 3' end, indicating that these nucleotides, in conjunction with the 3' terminus, are sufficient to recruit polymerase to the template. Northern blot and 5' rapid amplification of cDNA ends analysis of antigenome RNA indicated that antigenome initiation occurred at the first position of Le, irrespective of the terminal extension. This finding demonstrates that the 3' terminus of the RNA is not necessary for determining the antigenome initiation site. Data are presented which suggest that following recruitment to a promoter at the 3' end of Le, the polymerase is able to scan and respond to a promoter signal embedded within the RNA template.

* Corresponding author. Mailing address: Molecular and Cellular Pathology, Division of Pathology and Neuroscience, University of Dundee Medical School, Ninewells Hospital, Dundee, DD1 9SY United Kingdom. Fax: 44 1382 633952. Phone: 44 1382 496582. E-mail: r.fearns{at}dundee.ac.uk.

Journal of Virology, September 2005, p. 11311-11322, Vol. 79, No. 17
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.17.11311-11322.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Cordey, S., Roux, L. (2007). Further characterization of a paramyxovirus transcription initiation signal: search for required nucleotides upstream and importance of the N phase context. J. Gen. Virol. 88: 1555-1564 [Abstract] [Full Text]  
  • Tran, T.-L., Castagne, N., Bhella, D., Varela, P. F., Bernard, J., Chilmonczyk, S., Berkenkamp, S., Benhamo, V., Grznarova, K., Grosclaude, J., Nespoulos, C., Rey, F. A., Eleouet, J.-F. (2007). The nine C-terminal amino acids of the respiratory syncytial virus protein P are necessary and sufficient for binding to ribonucleoprotein complexes in which six ribonucleotides are contacted per N protein protomer. J. Gen. Virol. 88: 196-206 [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]  


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