Identification of influenza A nucleoprotein as an antiviral target

Influenza A remains a significant public health challenge because of the emergence of antigenically shifted or highly virulent strains. Antiviral resistance to available drugs such as adamantanes or neuraminidase inhibitors has appeared rapidly, creating a need for new antiviral targets and new drugs for influenza virus infections. Using forward chemical genetics, we have identified influenza A nucleoprotein (NP) as a druggable target and found a small-molecule compound, nucleozin, that triggers the aggregation of NP and inhibits its nuclear accumulation. Nucleozin impeded influenza A virus replication in vitro with a nanomolar median effective concentration (EC50) and protected mice challenged with lethal doses of avian influenza A H5N1. Our results demonstrate that viral NP is a valid target for the development of small-molecule therapies.

[1]  M. Peiris,et al.  Clinical features and rapid viral diagnosis of human disease associated with avian influenza A H5N1 virus , 1998, The Lancet.

[2]  Samson S. Y. Wong,et al.  Avian Influenza Virus Infections in Humans , 2006, Chest.

[3]  A. Lackenby,et al.  The potential impact of neuraminidase inhibitor resistant influenza , 2008, Current opinion in infectious diseases.

[4]  P. Digard,et al.  Edinburgh Research Explorer Identification of amino acid residues of influenza virus nucleoprotein essential for RNA binding , 2022 .

[5]  B. Stockwell.  Chemical genetics: ligand-based discovery of gene function , 2000, Nature Reviews Genetics.

[6]  R. Webster,et al.  H5N1 influenza--continuing evolution and spread. , 2006, The New England journal of medicine.

[7]  Lynnette Brammer,et al.  Infections with oseltamivir-resistant influenza A(H1N1) virus in the United States. , 2009, JAMA.

[8]  J. Davey,et al.  Identification of the sequence responsible for the nuclear accumulation of the influenza virus nucleoprotein in Xenopus oocytes , 1985, Cell.

[9]  M. Karplus,et al.  CHARMM: A program for macromolecular energy, minimization, and dynamics calculations , 1983 .

[10]  Wenjun Song,et al.  Nuclear Factor 90 Negatively Regulates Influenza Virus Replication by Interacting with Viral Nucleoprotein , 2009, Journal of Virology.

[11]  N. Pante,et al.  Nuclear import of influenza A viral ribonucleoprotein complexes is mediated by two nuclear localization sequences on viral nucleoprotein , 2007, Virology Journal.

[12]  C. Cambillau,et al.  Moth chemosensory protein exhibits drastic conformational changes and cooperativity on ligand binding , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[13]  E. Medcalf,et al.  Modulation of Nuclear Localization of the Influenza Virus Nucleoprotein through Interaction with Actin Filaments , 1999, Journal of Virology.

[14]  Herman Tse,et al.  Identification of Novel Small-Molecule Inhibitors of Severe Acute Respiratory Syndrome-Associated Coronavirus by Chemical Genetics , 2004, Chemistry & Biology.

[15]  K. Fujii,et al.  Contributions of Two Nuclear Localization Signals of Influenza A Virus Nucleoprotein to Viral Replication , 2006, Journal of Virology.

[16]  R. Krug,et al.  The primary function of RNA binding by the influenza A virus NS1 protein in infected cells: Inhibiting the 2'-5' oligo (A) synthetase/RNase L pathway. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[17]  David S. Goodsell,et al.  Automated docking using a Lamarckian genetic algorithm and an empirical binding free energy function , 1998, J. Comput. Chem..

[18]  S. Boulo,et al.  Nuclear traffic of influenza virus proteins and ribonucleoprotein complexes. , 2007, Virus research.

[19]  R. Webster,et al.  A DNA transfection system for generation of influenza A virus from eight plasmids. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[20]  P. Digard,et al.  The influenza virus nucleoprotein: a multifunctional RNA-binding protein pivotal to virus replication. , 2002, The Journal of general virology.

[21]  Sebastian Bonhoeffer,et al.  This PDF file includes: SOM Text , 2022 .

[22]  A. Moscona,et al.  Global transmission of oseltamivir-resistant influenza. , 2009, The New England journal of medicine.

[23]  K. Yuen,et al.  Characterization of SARS‐CoV main protease and identification of biologically active small molecule inhibitors using a continuous fluorescence‐based assay , 2004, FEBS Letters.

[24]  Samson S. Y. Wong,et al.  Delayed antiviral plus immunomodulator treatment still reduces mortality in mice infected by high inoculum of influenza A/H5N1 virus , 2008, Proceedings of the National Academy of Sciences.

[25]  R. Strausberg,et al.  From Knowing to Controlling: A Path from Genomics to Drugs Using Small Molecule Probes , 2003, Science.

[26]  A. Monto,et al.  Pandemic Influenza: An Inconvenient Mutation , 2009, Science.

[27]  R. Webster,et al.  Characterization of mutants of influenza A virus selected with the neuraminidase inhibitor 4-guanidino-Neu5Ac2en , 1996, Journal of virology.

[28]  R. Krug,et al.  The mechanism by which influenza A virus nucleoprotein forms oligomers and binds RNA , 2006, Nature.

[29]  M. Niepmann,et al.  Discontinuous native protein gel electrophoresis , 2006, Electrophoresis.