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Journal of Virology, October 1998, p. 8214-8219, Vol. 72, No. 10
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
The RNA Polymerase of Influenza Virus, Bound to the
5' End of Virion RNA, Acts in cis To Polyadenylate
mRNA
Leo L. M.
Poon,
David C.
Pritlove,
Jane
Sharps, and
George G.
Brownlee*
Sir William Dunn School of Pathology,
University of Oxford, Oxford, United Kingdom
Received 2 April 1998/Accepted 8 July 1998
We previously demonstrated, by limited mutagenesis, that conserved
sequence elements within the 5' end of influenza virus virion RNA
(vRNA) are required for the polyadenylation of mRNA in vitro. To
further characterize the nucleotide residues at the 5' end of vRNA
which might be involved in polyadenylation, a complete set of short and
long model vRNA-like templates with mutations at nucleotides 1' to 13'
(prime notation denotes numbering from the 5' end) of vRNA were
synthesized and transcribed in vitro. The products were assayed for
mRNA production with both reverse transcription-PCR and
[
-32P]ATP incorporation assays. Results from these
independent assays showed that vRNA templates with point mutations at
positions 2', 3', 7' to 9', and 11' to 13' synthesized polyadenylated
transcripts inefficiently compared with those with mutations at
positions 1', 4' to 6', and 10'. Positions 2', 3', 7' to 9', and 11'
are known to be involved in RNA polymerase binding. Furthermore,
residues at positions 11' to 13' are known to be involved in base
pairing between the 3' and 5' ends of vRNA. These findings demonstrate that the RNA polymerase has to bind to the 5' end of the template vRNA,
which must then interact with the 3' end of the same template for
polyadenylation to occur. These results support a model in which a
cis-acting RNA polymerase is required for the
polyadenylation of influenza virus.
*
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.
Journal of Virology, October 1998, p. 8214-8219, Vol. 72, No. 10
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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