Nucleoside Monophosphate Complex Structures of the Endonuclease Domain from the Influenza Virus Polymerase PA Subunit Reveal the Substrate Binding Site inside the Catalytic Center

被引:56
作者
Zhao, Cong [1 ]
Lou, Zhiyong [1 ]
Guo, Yu [2 ,3 ]
Ma, Ming [4 ]
Chen, Yutao [4 ]
Liang, Shuaiyi [1 ]
Zhang, Liang [2 ,3 ]
Chen, Shoudeng [4 ]
Li, Xuemei [4 ]
Liu, Yingfang [4 ]
Bartlam, Mark [2 ,3 ]
Rao, Zihe [1 ,2 ,3 ,4 ]
机构
[1] Tsinghua Univ, Struct Biol Lab, Beijing 100084, Peoples R China
[2] Nankai Univ, Coll Life Sci, Tianjin 300071, Peoples R China
[3] Nankai Univ, Tianjin State Lab Prot Sci, Tianjin 300071, Peoples R China
[4] Chinese Acad Sci, Inst Biophys, Natl Lab Macromol, Beijing 100101, Peoples R China
基金
中国国家自然科学基金;
关键词
RNA-POLYMERASE; CAP-BINDING; CRYSTAL-STRUCTURE; INHIBITORS; SOFTWARE; RESIDUES; MOTIFS; PB2;
D O I
10.1128/JVI.00911-09
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Highly pathogenic influenza virus strains currently in circulation pose a significant risk of a global pandemic. Following the reported crystal structure of the endonuclease domain from the avian influenza virus polymerase PA subunit, here we report the results of a systematic X-ray crystallographic analysis of its complex with adenosine, uridine, and thymidine nucleoside monophosphates (NMPs). Electron density corresponding to the monophosphate moiety of each nucleotide was apparent in each NMP complex and bound to the catalytic metal. A hydrophobic site was found to contribute to nucleoside binding. The NMP complex structures should represent the conformation of the bound product after nuclease cleavage. Moreover, one solvent molecule was found to occupy an equivalent position to the second reported Mn2+ ion, where it mediates the interaction between bound NMPs and the N-terminal PA domain in the presence of the Mg2+ ion. The results presented here indicate a possible cleavage mechanism and identify a distinct nucleotide binding pocket. The identification of this binding pocket opens a new avenue for anti-influenza drug discovery, targeting the cap-dependent endonuclease, in response to the worldwide threat of influenza.
引用
收藏
页码:9024 / 9030
页数:7
相关论文
共 23 条
[1]   PHENIX:: building new software for automated crystallographic structure determination [J].
Adams, PD ;
Grosse-Kunstleve, RW ;
Hung, LW ;
Ioerger, TR ;
McCoy, AJ ;
Moriarty, NW ;
Read, RJ ;
Sacchettini, JC ;
Sauter, NK ;
Terwilliger, TC .
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2002, 58 :1948-1954
[2]   MANIPULATION OF INTRACELLULAR MAGNESIUM CONTENT IN POLYMYXIN-B NONAPEPTIDE-SENSITIZED ESCHERICHIA-COLI BY IONOPHORE A23187 [J].
ALATOSSAVA, T ;
JUTTE, H ;
KUHN, A ;
KELLENBERGER, E .
JOURNAL OF BACTERIOLOGY, 1985, 162 (01) :413-419
[3]   MUTATIONAL ANALYSIS OF THE CONSERVED MOTIFS OF INFLUENZA-A VIRUS POLYMERASE BASIC-PROTEIN 1 [J].
BISWAS, SK ;
NAYAK, DP .
JOURNAL OF VIROLOGY, 1994, 68 (03) :1819-1826
[4]   Free R value: Cross-validation in crystallography [J].
Brunger, AT .
MACROMOLECULAR CRYSTALLOGRAPHY, PT B, 1997, 277 :366-396
[5]   Golgi manganese transport is required for rapamycin signaling in Saccharomyces cerevisiae [J].
Devasahayam, Gina ;
Burke, Daniel J. ;
Sturgill, Thomas W. .
GENETICS, 2007, 177 (01) :231-238
[6]   The cap-snatching endonuclease of influenza virus polymerase resides in the PA subunit [J].
Dias, Alexandre ;
Bouvier, Denis ;
Crepin, Thibaut ;
McCarthy, Andrew A. ;
Hart, Darren J. ;
Baudin, Florence ;
Cusack, Stephen ;
Ruigrok, Rob W. H. .
NATURE, 2009, 458 (7240) :914-918
[7]   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
[8]   Human infection with highly pathogenic H5N1 influenza virus [J].
Gambotto, Andrea ;
Barratt-Boyes, Simon M. ;
de Jong, Menno D. ;
Neumann, Gabriele ;
Kawaoka, Yoshihiro .
LANCET, 2008, 371 (9622) :1464-1475
[9]   The structural basis for cap binding by influenza virus polymerase subunit PB2 [J].
Guilligay, Delphine ;
Tarendeau, Franck ;
Resa-Infante, Patricia ;
Coloma, Rocio ;
Crepin, Thibaut ;
Sehr, Peter ;
Lewis, Joe ;
Ruigrok, Rob W. H. ;
Ortin, Juan ;
Hart, Darren J. ;
Cusack, Stephen .
NATURE STRUCTURAL & MOLECULAR BIOLOGY, 2008, 15 (05) :500-506
[10]   Mapping the domain structure of the influenza A virus polymerase acidic protein (PA) and its interaction with the basic protein 1 (PB1) subunit [J].
Guu, Tom S. Y. ;
Dong, Liping ;
Wittung-Stafshede, Pernilla ;
Tao, Yizhi J. .
VIROLOGY, 2008, 379 (01) :135-142