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 Pritlove, D. C.
Right arrow Articles by Brownlee, G. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Pritlove, D. C.
Right arrow Articles by Brownlee, G. G.

 Previous Article  |  Next Article 

J Virol, February 1998, p. 1280-1286, Vol. 72, No. 2
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Polyadenylation of Influenza Virus mRNA Transcribed In Vitro from Model Virion RNA Templates: Requirement for 5' Conserved Sequences

David C. Pritlove, Leo L. M. Poon, Ervin Fodor,dagger Jane Sharps, and George G. Brownlee*

Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom

Received 4 September 1997/Accepted 4 November 1997

Here we report the development of two independent assays which demonstrate for the first time that exogenous model RNA templates based on influenza virus virion RNA (vRNA) are transcribed in vitro to produce polyadenylated mRNA. We investigated the activities of mutated templates with known polymerase binding properties to test our model that polyadenylation occurs when a polymerase complex, which is bound to conserved 5' sequences of vRNA, prevents read-through of the U track at which polyadenylation subsequently occurs by reiterative copying. Mutated templates with perturbed polymerase binding sites (i.e., a deletion mutant lacking the first 4 5' residues and a Uright-arrowA point mutant at the third residue) initiated transcription in the in vitro assay but failed to produce polyadenylated transcripts, whereas an Aright-arrowU point mutant at the fourth residue, which retained polymerase binding properties similar to those of the wild type, produced polyadenylated transcripts. Our results show that nucleotides within the conserved 5' sequence are required for polyadenylation and support the hypothesis that polymerase binding to 5' sequences of the template is required for mRNA synthesis.


* Corresponding author. Mailing address: Chemical Pathology Unit, Sir William Dunn School of Pathology, University of Oxford, South Parks Rd., Oxford OX1 3RE, United Kingdom. Phone: (1865) 275559. Fax: (1865) 275556. E-mail: George.Brownlee{at}path.ox.ac.uk.

dagger Present address: Department of Microbiology, Mount Sinai School of Medicine, New York, NY 10029.




This article has been cited by other articles:

  • Vreede, F. T., Brownlee, G. G. (2007). Influenza Virion-Derived Viral Ribonucleoproteins Synthesize both mRNA and cRNA In Vitro. J. Virol. 81: 2196-2204 [Abstract] [Full Text]  
  • Barr, J. N., Rodgers, J. W., Wertz, G. W. (2005). The Bunyamwera Virus mRNA Transcription Signal Resides within both the 3' and the 5' Terminal Regions and Allows Ambisense Transcription from a Model RNA Segment. J. Virol. 79: 12602-12607 [Abstract] [Full Text]  
  • Barr, J. N., Wertz, G. W. (2005). Role of the Conserved Nucleotide Mismatch within 3'- and 5'-Terminal Regions of Bunyamwera Virus in Signaling Transcription. J. Virol. 79: 3586-3594 [Abstract] [Full Text]  
  • Gastaminza, P., Perales, B., Falcon, A. M., Ortin, J. (2003). Mutations in the N-Terminal Region of Influenza Virus PB2 Protein Affect Virus RNA Replication but Not Transcription. J. Virol. 77: 5098-5108 [Abstract] [Full Text]  
  • Lee, M.-K., Bae, S.-H., Park, C.-J., Cheong, H.-K., Cheong, C., Choi, B.-S. (2003). A single-nucleotide natural variation (U4 to C4) in an influenza A virus promoter exhibits a large structural change: implications for differential viral RNA synthesis by RNA-dependent RNA polymerase. Nucleic Acids Res 31: 1216-1223 [Abstract] [Full Text]  
  • Honda, A., Mizumoto, K., Ishihama, A. (2002). Minimum molecular architectures for transcription and replication of the influenza virus. Proc. Natl. Acad. Sci. USA 99: 13166-13171 [Abstract] [Full Text]  
  • Brownlee, G. G., Sharps, J. L. (2002). The RNA Polymerase of Influenza A Virus Is Stabilized by Interaction with Its Viral RNA Promoter. J. Virol. 76: 7103-7113 [Abstract] [Full Text]  
  • Portela, A., Digard, P. (2002). The influenza virus nucleoprotein: a multifunctional RNA-binding protein pivotal to virus replication. J. Gen. Virol. 83: 723-734 [Abstract] [Full Text]  
  • Bae, S.-H., Cheong, H.-K., Lee, J.-H., Cheong, C., Kainosho, M., Choi, B.-S. (2001). Structural features of an influenza virus promoter and their implications for viral RNA synthesis. Proc. Natl. Acad. Sci. USA 98: 10602-10607 [Abstract] [Full Text]  
  • Leahy, M. B., Dobbyn, H. C., Brownlee, G. G. (2001). Hairpin Loop Structure in the 3' Arm of the Influenza A Virus Virion RNA Promoter Is Required for Endonuclease Activity. J. Virol. 75: 7042-7049 [Abstract] [Full Text]  
  • Hooker, L., Strong, R., Adams, R., Handa, B., Merrett, J. H., Martin, J. A., Klumpp, K. (2001). A sensitive, single-tube assay to measure the enzymatic activities of influenza RNA polymerase and other poly(A) polymerases: application to kinetic and inhibitor analysis. Nucleic Acids Res 29: 2691-2698 [Abstract] [Full Text]  
  • Crescenzo-Chaigne, B., van der Werf, S. (2001). Nucleotides at the extremities of the viral RNA of influenza C virus are involved in type-specific interactions with the polymerase complex. J. Gen. Virol. 82: 1075-1083 [Abstract] [Full Text]  
  • Leahy, M. B., Pritlove, D. C., Poon, L. L. M., Brownlee, G. G. (2001). Mutagenic Analysis of the 5' Arm of the Influenza A Virus Virion RNA Promoter Defines the Sequence Requirements for Endonuclease Activity. J. Virol. 75: 134-142 [Abstract] [Full Text]  
  • Aragón, T., de la Luna, S., Novoa, I., Carrasco, L., Ortín, J., Nieto, A. (2000). Eukaryotic Translation Initiation Factor 4GI Is a Cellular Target for NS1 Protein, a Translational Activator of Influenza Virus. Mol. Cell. Biol. 20: 6259-6268 [Abstract] [Full Text]  
  • Greger, I. H., Aranda, A., Proudfoot, N. (2000). Balancing transcriptional interference and initiation on the GAL7 promoter of Saccharomycescerevisiae. Proc. Natl. Acad. Sci. USA 10.1073/pnas.140217697v1 [Abstract] [Full Text]  
  • Perales, B., Sanz-Ezquerro, J. J., Gastaminza, P., Ortega, J., Santarén, J. F., Ortín, J., Nieto, A. (2000). The Replication Activity of Influenza Virus Polymerase Is Linked to the Capacity of the PA Subunit To Induce Proteolysis. J. Virol. 74: 1307-1312 [Abstract] [Full Text]  
  • Ortega, J., Martín-Benito, J., Zürcher, T., Valpuesta, J. M., Carrascosa, J. L., Ortín, J. (2000). Ultrastructural and Functional Analyses of Recombinant Influenza Virus Ribonucleoproteins Suggest Dimerization of Nucleoprotein during Virus Amplification. J. Virol. 74: 156-163 [Abstract] [Full Text]  
  • Poon, L. L. M., Fodor, E., Brownlee, G. G. (2000). Polyuridylated mRNA Synthesized by a Recombinant Influenza Virus Is Defective in Nuclear Export. J. Virol. 74: 418-427 [Abstract] [Full Text]  
  • Medcalf, L., Poole, E., Elton, D., Digard, P. (1999). Temperature-Sensitive Lesions in Two Influenza A Viruses Defective for Replicative Transcription Disrupt RNA Binding by the Nucleoprotein. J. Virol. 73: 7349-7356 [Abstract] [Full Text]  
  • Zheng, H., Lee, H. A., Palese, P., García-Sastre, A. (1999). Influenza A Virus RNA Polymerase Has the Ability To Stutter at the Polyadenylation Site of a Viral RNA Template during RNA Replication. J. Virol. 73: 5240-5243 [Abstract] [Full Text]  
  • Poon, L. L. M., Pritlove, D. C., Fodor, E., Brownlee, G. G. (1999). Direct Evidence that the Poly(A) Tail of Influenza A Virus mRNA Is Synthesized by Reiterative Copying of a U Track in the Virion RNA Template. J. Virol. 73: 3473-3476 [Abstract] [Full Text]  
  • Pritlove, D. C., Poon, L. L. M., Devenish, L. J., Leahy, M. B., Brownlee, G. G. (1999). A Hairpin Loop at the 5' End of Influenza A Virus Virion RNA Is Required for Synthesis of Poly(A)+ mRNA In Vitro. J. Virol. 73: 2109-2114 [Abstract] [Full Text]  
  • Mena, I., Jambrina, E., Albo, C., Perales, B., Ortín, J., Arrese, M., Vallejo, D., Portela, A. (1999). Mutational Analysis of Influenza A Virus Nucleoprotein: Identification of Mutations That Affect RNA Replication. J. Virol. 73: 1186-1194 [Abstract] [Full Text]  
  • Gonzalez, S., Ortin, J. (1999). Characterization of Influenza Virus PB1 Protein Binding to Viral RNA: Two Separate Regions of the Protein Contribute to the Interaction Domain. J. Virol. 73: 631-637 [Abstract] [Full Text]  
  • Poon, L. L. M., Pritlove, D. C., Sharps, J., Brownlee, G. G. (1998). The RNA Polymerase of Influenza Virus, Bound to the 5' End of Virion RNA, Acts in cis To Polyadenylate mRNA. J. Virol. 72: 8214-8219 [Abstract] [Full Text]  
  • Fodor, E., Palese, P., Brownlee, G. G., Garcia-Sastre, A. (1998). Attenuation of Influenza A Virus mRNA Levels by Promoter Mutations. J. Virol. 72: 6283-6290 [Abstract] [Full Text]  
  • Honda, A., Endo, A., Mizumoto, K., Ishihama, A. (2001). Differential Roles of Viral RNA and cRNA in Functional Modulation of the Influenza Virus RNA Polymerase. J. Biol. Chem. 276: 31179-31185 [Abstract] [Full Text]  
  • Greger, I. H., Aranda, A., Proudfoot, N. (2000). Balancing transcriptional interference and initiation on the GAL7 promoter of Saccharomycescerevisiae. Proc. Natl. Acad. Sci. USA 97: 8415-8420 [Abstract] [Full Text]