Previous Article | Next Article ![]()
J Virol, February 1998, p. 1280-1286, Vol. 72, No. 2
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 U
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

A point
mutant at the third residue) initiated transcription in the in vitro
assay but failed to produce polyadenylated transcripts, whereas an
A
U 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.
Present address: Department of Microbiology, Mount Sinai School of
Medicine, New York, NY 10029.
This article has been cited by other articles:
| J. Bacteriol. | Mol. Cell. Biol. | Microbiol. Mol. Biol. Rev. |
|---|
| Clin. Vaccine Immunol. | ALL ASM JOURNALS |
|---|