Structure and nuclear import function of the C-terminal domain of influenza virus polymerase PB2 subunit

被引:263
作者
Tarendeau, Franck
Boudet, Julien
Guilligay, Delphine
Mas, Philippe J.
Bougault, Catherine M.
Boulo, Sebastien
Baudin, Florence
Ruigrok, Rob W. H.
Daigle, Nathalie
Ellenberg, Jan
Cusack, Stephen
Simorre, Jean-Pierre
Hart, Darren J.
机构
[1] European Mol Biol Lab, F-38042 Grenoble, France
[2] UJF, UMR CNRS CEA 5075, Inst Biol Struct Jean Pierre Ebel, F-38027 Grenoble 1, France
[3] UJF, UMR CNRS 5233, EMB, Unit Virus Host Cell Interact, F-38042 Grenoble 9, France
关键词
D O I
10.1038/nsmb1212
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The trimeric influenza virus polymerase, comprising subunits PA, PB1 and PB2, is responsible for transcription and replication of the segmented viral RNA genome. Using a novel library-based screening technique called expression of soluble proteins by random incremental truncation (ESPRIT), we identified an independently folded C-terminal domain from PB2 and determined its solution structure by NMR. Using green fluorescent protein fusions, we show that both the domain and the full-length PB2 subunit are efficiently imported into the nucleus dependent on a previously overlooked bipartite nuclear localization sequence (NLS). The crystal structure of the domain complexed with human importin alpha 5 shows how the last 20 residues unfold to permit binding to the import factor. The domain contains three surface residues implicated in adaptation from avian to mammalian hosts. One of these tethers the NLS-containing peptide to the core of the domain in the unbound state.
引用
收藏
页码:229 / 233
页数:5
相关论文
共 33 条
[1]   3D structure of the influenza virus polymerase complex:: Localization of subunit domains [J].
Area, E ;
Martín-Benito, J ;
Gastaminza, P ;
Torreira, E ;
Valpuesta, JM ;
Carrascosa, JL ;
Ortín, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (01) :308-313
[2]   A minimal peptide substrate in biotin holoenzyme synthetase-catalyzed biotinylation [J].
Beckett, D ;
Kovaleva, E ;
Schatz, PJ .
PROTEIN SCIENCE, 1999, 8 (04) :921-929
[3]   Crystallography & NMR system:: A new software suite for macromolecular structure determination [J].
Brunger, AT ;
Adams, PD ;
Clore, GM ;
DeLano, WL ;
Gros, P ;
Grosse-Kunstleve, RW ;
Jiang, JS ;
Kuszewski, J ;
Nilges, M ;
Pannu, NS ;
Read, RJ ;
Rice, LM ;
Simonson, T ;
Warren, GL .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1998, 54 :905-921
[4]   Crystallographic analysis of the specific yet versatile recognition of distinct nuclear localization signals by karyopherin α [J].
Conti, E ;
Kuriyan, J .
STRUCTURE, 2000, 8 (03) :329-338
[5]   Fatal outcome of human influenza A (H5N1) is associated with high viral load and hypercytokinemia [J].
de Jong, Menno D. ;
Simmons, Cameron P. ;
Thanh, Tran Tan ;
Hien, Vo Minh ;
Smith, Gavin J. D. ;
Chau, Tran Nguyen Bich ;
Hoang, Dang Minh ;
Chau, Nguyen Van Vinh ;
Khanh, Truong Huu ;
Dong, Vo Cong ;
Qui, Phan Tu ;
Van Cam, Bach ;
Ha, Do Quang ;
Guan, Yi ;
Peiris, J. S. Malik ;
Chinh, Nguyen Tran ;
Hien, Tran Tinh ;
Farrar, Jeremy .
NATURE MEDICINE, 2006, 12 (10) :1203-1207
[6]   MOLECULAR-CLONING AND SEQUENCING OF INFLUENZA-VIRUS A/VICTORIA/3/75 POLYMERASE GENES - SEQUENCE EVOLUTION AND PREDICTION OF POSSIBLE FUNCTIONAL DOMAINS [J].
DELALUNA, S ;
MARTINEZ, C ;
ORTIN, J .
VIRUS RESEARCH, 1989, 13 (02) :143-156
[7]   Role of ran binding protein 5 in nuclear import and assembly of the influenza virus RNA polymerase complex [J].
Deng, Tao ;
Engelhardt, Othmar G. ;
Thomas, Benjamin ;
Akoulitchev, Alexandre V. ;
Brownlee, George G. ;
Fodor, Ervin .
JOURNAL OF VIROLOGY, 2006, 80 (24) :11911-11919
[8]   Two aromatic residues in the PB2 subunit of influenza A RNA polymerase are crucial for cap binding [J].
Fechter, P ;
Mingay, L ;
Sharps, J ;
Chambers, A ;
Fodor, E ;
Brownlee, GG .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (22) :20381-20388
[9]   The PA subunit is required for efficient nuclear accumulation of the PB1 subunit of the influenza A virus RNA polymerase complex [J].
Fodor, E ;
Smith, M .
JOURNAL OF VIROLOGY, 2004, 78 (17) :9144-9153
[10]   Structural basis for the specificity of bipartite nuclear localization sequence binding by importin-α [J].
Fontes, MRM ;
Teh, T ;
Jans, D ;
Brinkworth, RI ;
Kobe, B .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (30) :27981-27987