Nucleoside monophosphate complex structures of the endonuclease domain from the influenza virus polymerase PA subunit reveal the substrate binding site inside the catalytic center.
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 Mn(2+) ion, where it mediates the interaction between bound NMPs and the N-terminal PA domain in the presence of the Mg(2+) 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.
Zhao
Cong
C
Laboratory of Structural Biology, Room 201, New Life Sciences Building, Tsinghua University, Beijing 100084, China.
Lou
Zhiyong
Z
Guo
Yu
Y
Ma
Ming
M
Chen
Yutao
Y
Liang
Shuaiyi
S
Zhang
Liang
L
Chen
Shoudeng
S
Li
Xuemei
X
Liu
Yingfang
Y
Bartlam
Mark
M
Rao
Zihe
Z
D001665:Binding Sites; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D004720:Endonucleases / Q000737:chemistry*; D053124:Influenza A Virus, H5N1 Subtype / Q000201:enzymology*; D008274:Magnesium / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D009711:Nucleotides / Q000737:chemistry* / Q000378:metabolism; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry* / Q000378:metabolism; D012997:Solvents; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism
D009711:Nucleotides; C061290:PA protein, influenza viruses; D012997:Solvents; D014764:Viral Proteins; D008345:Manganese; D012324:RNA-Dependent RNA Polymerase; D004720:Endonucleases; D008274:Magnesium