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Journal of Virology, February 2001, p. 1899-1908, Vol. 75, No. 4
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.4.1899-1908.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Cellular Splicing Factor RAF-2p48/NPI-5/BAT1/UAP56 Interacts
with the Influenza Virus Nucleoprotein and Enhances Viral
RNA Synthesis
Fumitaka
Momose,1,
Christopher F.
Basler,2
Robert E.
O'Neill,2,
Akihiro
Iwamatsu,3
Peter
Palese,2 and
Kyosuke
Nagata1,*
Laboratory of Molecular Medical Engineering,
Department of Biological Information, Graduate School of Bioscience and
Biotechnology, Tokyo Institute of Technology, Midori-ku, Yokohama
226-8501,1 and Central Laboratories
for Key Technology, Kirin Brewery Company, Kanazawa-ku, Yokohama,
Kanagawa 236-0004,3 Japan, and
Department of Microbiology, Mount Sinai School of Medicine,
New York, New York 10029-65742
Received 2 August 2000/Accepted 16 November 2000
Previous biochemical data identified a host cell fraction,
designated RAF-2, which stimulated influenza virus RNA synthesis. A
48-kDa polypeptide (RAF-2p48), a cellular splicing factor belonging to
the DEAD-box family of RNA-dependent ATPases previously designated BAT1
(also UAP56), has now been identified as essential for RAF-2 stimulatory activity. Additionally, RAF-2p48 was independently identified as an influenza virus nucleoprotein (NP)-interacting protein, NPI-5, in a yeast two-hybrid screen of a mammalian cDNA library. In vitro, RAF-2p48 interacted with free NP but not with NP
bound to RNA, and the RAF-2p48-NP complex was dissociated following addition of free RNA. Furthermore, RAF-2p48 facilitated formation of
the NP-RNA complexes that likely serve as templates for the viral RNA
polymerase. RAF-2p48 was shown, in both in vitro binding assays and the
yeast two-hybrid system, to bind to the amino-terminal region of NP, a
domain essential for RNA binding. Together, these observations suggest
that RAF-2p48 facilitates NP-RNA interaction, thus leading to enhanced
influenza virus RNA synthesis.
*
Corresponding author. Mailing address: Laboratory of
Molecular Medical Engineering, Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of
Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan. Phone:
81 45-924-5798. Fax: 81 45-924-5804. E-mail:
knagata{at}bio.titech.ac.jp.

Present address: Department of Virology, Center for Basic Research,
The Kitasato Institute, Minato-ku, Tokyo 108-8642,
Japan.

Present address: Department of Viral Vaccine Research,
Wyeth-Lederle Vaccines and Pediatrics, Pearl River, NY
10965.
Journal of Virology, February 2001, p. 1899-1908, Vol. 75, No. 4
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.4.1899-1908.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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