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Journal of Virology, August 2001, p. 7042-7049, Vol. 75, No. 15
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.15.7042-7049.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Hairpin Loop Structure in the 3' Arm of the
Influenza A Virus Virion RNA Promoter Is Required for
Endonuclease Activity
Michael B.
Leahy,
Helen C.
Dobbyn,
and
George G.
Brownlee*
Sir William Dunn School of Pathology,
University of Oxford, Oxford, United Kingdom
Received 13 February 2001/Accepted 25 April 2001
Previous studies have shown that the 5' arm of the influenza A
virus virion RNA promoter requires a hairpin loop structure for
efficient endonuclease activity of influenza virus RNA polymerase, an
activity that is required for the cap-snatching activity of primers
from host pre-mRNA. Here we examine whether a hairpin loop is also
required in the 3' arm of the viral RNA promoter. We study point
mutations at each nucleotide position (1 to 12) within the 3' arm of
the promoter as well as complementary "rescue" mutations which
restored base pairing in the stem of a potential hairpin loop. Our
results suggest that endonuclease activity is absolutely dependent on
the presence of a 3' hairpin loop structure. This is the first direct
evidence for RNA secondary structure within the 3' arm being required
for a specific stage, i.e., endonuclease cleavage, in the influenza
virus replicative cycle.
*
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: 44 1865 275559. Fax: 44 1865 275556. E-mail:
George.Brownlee{at}path.ox.ac.uk.

Present address: School of Biological Sciences, Biochemistry
Division, University of Manchester, Manchester, United
Kingdom.
Journal of Virology, August 2001, p. 7042-7049, Vol. 75, No. 15
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.15.7042-7049.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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