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Journal of Virology, September 1999, p. 7349-7356, Vol. 73, No. 9
Division of Virology, Department of
Pathology, University of Cambridge, Cambridge CB2 1QP, United
Kingdom
Received 11 March 1999/Accepted 25 May 1999
The negative-sense segmented RNA genome of influenza virus is
transcribed into capped and polyadenylated mRNAs, as well as full-length replicative intermediates (cRNAs). The mechanism that regulates the two forms of transcription remains unclear,
although several lines of evidence imply a role for the viral
nucleoprotein (NP). In particular, temperature-shift and
biochemical analyses of the temperature-sensitive viruses A/WSN/33
ts56 and A/FPV/Rostock/34/Giessen tsG81
containing point mutations within the NP coding region have indicated
specific defects in replicative transcription at the nonpermissive
temperature. To identify the functional defect, we introduced the
relevant mutations into the NP of influenza virus strain A/PR/8/34.
Both mutants were temperature sensitive for influenza virus gene
expression in transient-transfection experiments but localized and
accumulated normally in transfected cells. Similarly, the mutants
retained the ability to self-associate and interact with the virus
polymerase complex whether synthesized at the permissive or the
nonpermissive temperatures. In contrast, the mutant NPs were
defective for RNA binding when expressed at the nonpermissive
temperature but not when expressed at 30°C. This suggests that the
RNA-binding activity of NP is required for replicative transcription.
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Temperature-Sensitive Lesions in Two Influenza A Viruses
Defective for Replicative Transcription Disrupt RNA Binding by
the Nucleoprotein
*
Corresponding author. Mailing address: Division of
Virology, Department of Pathology, University of Cambridge, Tennis
Court Rd., Cambridge CB2 1QP, United Kingdom. Phone: 44-1223-336918. Fax: 44-1223-336926. E-mail:
pd1{at}mole.bio.cam.ac.uk.
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