REPLICATION OF INFLUENZA VIRUS

Summary

Principal Investigator: ROBERT KRUG
Affiliation: University of Texas
Country: USA
Abstract: DESCRIPTION (provided by applicant): The biological activities of the NS1 protein of influenza A virus (NS1A protein) differ markedly from those of the NS1 protein of influenza B virus (NS1B protein). The NS1 proteins of these two genera of influenza viruses, which cause widespread human disease, likely mediate many of the viral countermeasures that combat cellular antiviral responses. Our overall aim is to identify all the activities of the NS1A and NS1B proteins and to elucidate how these different activities counter the cellular antiviral response and/or have other functions during virus replication. At early times after infection with either influenza A or B virus the transcription of cellular genes controlled by IFN (interferon)-stimulated response elements (ISREs) is activated without the involvement of IFNalpha/beta. Mature cellular mRNAs are produced in influenza B-, but not in influenza A-, infected cells. Our aims are to determine: (1) whether the absence of mature cellular mRNAs in influenza A virus-infected cells is caused by the block in 3' end processing of cellular pre-mRNA that is mediated by the NS1A protein, a function lacked by the NS1B protein; and (2) the mechanism by which ISREs of these cellular genes are activated in influenza A and B virus-infected cells. The NS1A protein contains a nuclear export signal (NES) which is activated between 2 and 4 hours post-infection. Our aim is to determine the mechanism of activation of this NES and its role during influenza A virus infection. The NS1B protein, but not the NS1A protein, binds ISG15, a human ubiquitin-like protein that is induced by either IFNalpha/beta or virus infection, and inhibits the conjugation of ISG15 to its target proteins. Because these protein targets have not yet been identified, it is not known why ISG15 conjugation would be detrimental to influenza B virus infection. Our goals are: (i) to establish how the conjugation of ISG15 to its target protein affects influenza B virus replication; and (ii) to identify the protein targets of ISG15 conjugation and to determine how this conjugation affects the activities of these target proteins. Other cellular proteins also likely bind to the NS1A and/or NS1B protein, e.g., the SRp54 protein that binds to NS1A, but not to NS1B, and our aim is to determine whether the binding of these other proteins has a functional role in influenza virus-infected cells. These functional analyzes of the NS1A and NS1B proteins will be complemented by structural studies using both nuclear magnetic resonance and X-ray crystallography.
Funding Period: 1977-07-01 - 2007-12-14
more information: NIH RePORT

Top Publications

  1. pmc Cellular DDX21 RNA Helicase Inhibits Influenza A Virus Replication but Is Counteracted by the Viral NS1 Protein
    Guifang Chen
    Department of Molecular Biosciences, Center of Infectious Disease, Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX 78712, USA
    Cell Host Microbe 15:484-93. 2014
  2. pmc Reciprocal inhibition between intracellular antiviral signaling and the RNAi machinery in mammalian cells
    Gil Ju Seo
    The University of Texas at Austin, Molecular Genetics and Microbiology, 1 University Station A5000, Austin, TX 78712 0162, USA
    Cell Host Microbe 14:435-45. 2013
  3. pmc Interferon-induced ISG15 pathway: an ongoing virus-host battle
    Chen Zhao
    Department of Molecular Genetics and Microbiology, Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX 78712, USA
    Trends Microbiol 21:181-6. 2013
  4. pmc The ESEV PDZ-binding motif of the avian influenza A virus NS1 protein protects infected cells from apoptosis by directly targeting Scribble
    Hongbing Liu
    Department of Molecular Biology and Microbiology, Baylor College of Medicine, BCMD 824DB, Houston, TX 77030, USA
    J Virol 84:11164-74. 2010
  5. pmc Species specificity of the NS1 protein of influenza B virus: NS1 binds only human and non-human primate ubiquitin-like ISG15 proteins
    Haripriya Sridharan
    Institute for Cellular and Molecular Biology, Section of Molecular Genetics and Microbiology, University of Texas, Austin, Texas 78712, USA
    J Biol Chem 285:7852-6. 2010
  6. pmc Structures of influenza A proteins and insights into antiviral drug targets
    Kalyan Das
    Center for Advanced Biotechnology and Medicine, Piscataway, New Jersey, USA
    Nat Struct Mol Biol 17:530-8. 2010
  7. pmc Influenza A virus strains that circulate in humans differ in the ability of their NS1 proteins to block the activation of IRF3 and interferon-β transcription
    Rei Lin Kuo
    Institute for Cellular and Molecular Biology, Section of Molecular Genetics and Microbiology, University of Texas at Austin, Austin, TX 78746, USA
    Virology 408:146-58. 2010
  8. pmc Backbone and Ile-δ1, Leu, Val methyl 1H, 13C and 15N NMR chemical shift assignments for human interferon-stimulated gene 15 protein
    Cuifeng Yin
    Department of Molecular Biology and Biochemistry, Center for Advanced Biotechnology and Medicine, Northeast Structural Genomics Consortium, Rutgers, The State University of New Jersey, Piscataway, NJ 08854, USA
    Biomol NMR Assign 5:215-9. 2011
  9. pmc The virulence of 1997 H5N1 influenza viruses in the mouse model is increased by correcting a defect in their NS1 proteins
    April Spesock
    Centers for Disease Control and Prevention, Influenza Division, 1600 Clifton Road NE, MD G16, Atlanta, GA 30333, USA
    J Virol 85:7048-58. 2011
  10. pmc Dimer interface of the effector domain of non-structural protein 1 from influenza A virus: an interface with multiple functions
    James M Aramini
    Center for Advanced Biotechnology and Medicine, Northeast Structural Genomics Consortium, Department of Molecular Biology and Biochemistry, Rutgers, The State University of New Jersey, Piscataway, New Jersey 08854, USA
    J Biol Chem 286:26050-60. 2011

Scientific Experts

  • Kalyan Das
  • James M Aramini
  • ROBERT KRUG
  • Krister Melen
  • Chen Zhao
  • Rei Lin Kuo
  • Tien Ying Hsiang
  • Ji Young Min
  • Cuifeng Yin
  • Karen Y Twu
  • Diana L Noah
  • Haripriya Sridharan
  • Gaetano T Montelione
  • Shoudong Li
  • Guifang Chen
  • Gil Ju Seo
  • Li Chung Ma
  • April Spesock
  • Rongjin Guan
  • Meghana Malur
  • Hongbing Liu
  • G V T Swapna
  • Laura L Newcomb
  • Ganes C Sen
  • Anahita Dastur
  • Jukka Sirén
  • Jon M Huibregtse
  • Chien Hung Liu
  • Ligang Zhou
  • Justin M Pare
  • Mark N Collins
  • Jennifer E Cox
  • Christopher S Sullivan
  • James M Burke
  • Rodney P Kincaid
  • Teva Phanaksri
  • Paul G Leonard
  • Li Mei Chen
  • Brendan R Amer
  • Charles T Davis
  • Ruben O Donis
  • Ian A York
  • John R Cort
  • Aleksandr S Lipatov
  • Bradley L Njaa
  • M Jaber Hossain
  • Ronald T Javier
  • Eugene C Dow
  • Lisa Golebiewski
  • Andrew P Rice
  • Yizhi Jane Tao
  • Yunyun Jiang
  • Qiaozhen Ye
  • Jesper Marklund
  • Javed A Khan
  • Asli Ertekin
  • Liang Tong
  • Ilkka Julkunen
  • Paul B Fisher
  • Sampsa Matikainen
  • Devanand Sarkar
  • Taija Pietilä
  • Sylvie Beaudenon
  • Ping Rao
  • Tadaatsu Imaizumi
  • Melissa Kelley
  • John Hiscott
  • Rongtuan Lin
  • Carilee Denison
  • Steven Gygi

Detail Information

Publications27

  1. pmc Cellular DDX21 RNA Helicase Inhibits Influenza A Virus Replication but Is Counteracted by the Viral NS1 Protein
    Guifang Chen
    Department of Molecular Biosciences, Center of Infectious Disease, Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX 78712, USA
    Cell Host Microbe 15:484-93. 2014
    ....
  2. pmc Reciprocal inhibition between intracellular antiviral signaling and the RNAi machinery in mammalian cells
    Gil Ju Seo
    The University of Texas at Austin, Molecular Genetics and Microbiology, 1 University Station A5000, Austin, TX 78712 0162, USA
    Cell Host Microbe 14:435-45. 2013
    ..Our results suggest that, unlike in lower eukaryotes, mammalian RISC is not antiviral in some contexts, but rather RISC has been co-opted to negatively regulate toxic host antiviral effectors via microRNAs. ..
  3. pmc Interferon-induced ISG15 pathway: an ongoing virus-host battle
    Chen Zhao
    Department of Molecular Genetics and Microbiology, Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX 78712, USA
    Trends Microbiol 21:181-6. 2013
    ..Only the antiviral activity of human and non-human primate ISG15s can be blocked, providing one possible explanation for the restriction of influenza B virus to humans...
  4. pmc The ESEV PDZ-binding motif of the avian influenza A virus NS1 protein protects infected cells from apoptosis by directly targeting Scribble
    Hongbing Liu
    Department of Molecular Biology and Microbiology, Baylor College of Medicine, BCMD 824DB, Houston, TX 77030, USA
    J Virol 84:11164-74. 2010
    ..This study shows that one function of the avian ESEV PBM is to reduce apoptosis during infection through disruption of Scribble's proapoptotic function...
  5. pmc Species specificity of the NS1 protein of influenza B virus: NS1 binds only human and non-human primate ubiquitin-like ISG15 proteins
    Haripriya Sridharan
    Institute for Cellular and Molecular Biology, Section of Molecular Genetics and Microbiology, University of Texas, Austin, Texas 78712, USA
    J Biol Chem 285:7852-6. 2010
    ..Consequently, the ISG15 proteins of humans and non-human primates are the only mammalian ISG15 proteins that would bind NS1B...
  6. pmc Structures of influenza A proteins and insights into antiviral drug targets
    Kalyan Das
    Center for Advanced Biotechnology and Medicine, Piscataway, New Jersey, USA
    Nat Struct Mol Biol 17:530-8. 2010
    ..The structures of domains of the influenza RNA-dependent RNA polymerase and the nonstructural NS1A protein provide opportunities for targeting these proteins to inhibit viral replication...
  7. pmc Influenza A virus strains that circulate in humans differ in the ability of their NS1 proteins to block the activation of IRF3 and interferon-β transcription
    Rei Lin Kuo
    Institute for Cellular and Molecular Biology, Section of Molecular Genetics and Microbiology, University of Texas at Austin, Austin, TX 78746, USA
    Virology 408:146-58. 2010
    ..Further, we show that TRIM25 binds the NS1 protein whether or not IRF3 activation is blocked, demonstrating that binding of TRIM25 by the NS1 protein does not necessarily lead to the blocking of IRF3 activation...
  8. pmc Backbone and Ile-δ1, Leu, Val methyl 1H, 13C and 15N NMR chemical shift assignments for human interferon-stimulated gene 15 protein
    Cuifeng Yin
    Department of Molecular Biology and Biochemistry, Center for Advanced Biotechnology and Medicine, Northeast Structural Genomics Consortium, Rutgers, The State University of New Jersey, Piscataway, NJ 08854, USA
    Biomol NMR Assign 5:215-9. 2011
    ..These resonance assignments provide the basis for future structural and functional solution NMR studies of the biologically important human ISG15 protein...
  9. pmc The virulence of 1997 H5N1 influenza viruses in the mouse model is increased by correcting a defect in their NS1 proteins
    April Spesock
    Centers for Disease Control and Prevention, Influenza Division, 1600 Clifton Road NE, MD G16, Atlanta, GA 30333, USA
    J Virol 85:7048-58. 2011
    ..We conclude that strengthening CPSF30 binding by the NS1 protein of 1997 H5N1 viruses enhances virulence in mice by increasing the systemic spread of the virus from the lungs, particularly to the brain...
  10. pmc Dimer interface of the effector domain of non-structural protein 1 from influenza A virus: an interface with multiple functions
    James M Aramini
    Center for Advanced Biotechnology and Medicine, Northeast Structural Genomics Consortium, Department of Molecular Biology and Biochemistry, Rutgers, The State University of New Jersey, Piscataway, New Jersey 08854, USA
    J Biol Chem 286:26050-60. 2011
    ....
  11. pmc Structural basis for the sequence-specific recognition of human ISG15 by the NS1 protein of influenza B virus
    Rongjin Guan
    Center for Advanced Biotechnology and Medicine, Rutgers University, Piscataway NJ 08854, USA
    Proc Natl Acad Sci U S A 108:13468-73. 2011
    ..Nonetheless, transfection assays show that NS1B-NTR binding of ISG15 is responsible for the inhibition of interferon-induced ISG15 conjugation in cells...
  12. pmc ISG15 conjugation system targets the viral NS1 protein in influenza A virus-infected cells
    Chen Zhao
    Institute for Cellular and Molecular Biology, Section of Molecular Genetics and Microbiology, University of Texas at Austin, TX 78712, USA
    Proc Natl Acad Sci U S A 107:2253-8. 2010
    ....
  13. pmc Interferon-induced ISG15 conjugation inhibits influenza A virus gene expression and replication in human cells
    Tien Ying Hsiang
    Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX 78712, USA
    J Virol 83:5971-7. 2009
    ..In contrast, we show that in mouse embryo fibroblasts ISG15 conjugation not only does not affect influenza A virus replication but also does not contribute to the IFN-induced antiviral activity against influenza A virus gene expression...
  14. pmc Influenza a virus polymerase is an integral component of the CPSF30-NS1A protein complex in infected cells
    Rei Lin Kuo
    Institute for Cellular and Molecular Biology, University of Texas at Austin, 78712, USA
    J Virol 83:1611-6. 2009
    ....
  15. pmc Interaction of the influenza a virus nucleocapsid protein with the viral RNA polymerase potentiates unprimed viral RNA replication
    Laura L Newcomb
    Institute for Cellular and Molecular Biology, University of Texas at Austin, 2500 Speedway, Austin, TX 78712, USA
    J Virol 83:29-36. 2009
    ....
  16. ncbi Influenza virus virulence and its molecular determinants
    Diana L Noah
    Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, Texas 78712, USA
    Adv Virus Res 65:121-45. 2005
  17. ncbi Herc5, an interferon-induced HECT E3 enzyme, is required for conjugation of ISG15 in human cells
    Anahita Dastur
    Institute for Cellular and Molecular Biology, Section of Molecular Genetics and Microbiology, The University of Texas at Austin, 78712, USA
    J Biol Chem 281:4334-8. 2006
    ..These results demonstrate that Herc5 is required for conjugation of ISG15 to a broad spectrum of target proteins in human cells...
  18. ncbi Binding of the influenza A virus NS1 protein to PKR mediates the inhibition of its activation by either PACT or double-stranded RNA
    Shoudong Li
    Department of Molecular Genetics, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH 44195, USA
    Virology 349:13-21. 2006
    ..In addition, we demonstrated that PKR activation is inhibited in cells infected with a recombinant influenza A virus expressing NS1A mutant protein that cannot bind RNA, as is the case in cells infected with wild-type influenza A virus...
  19. pmc The CPSF30 binding site on the NS1A protein of influenza A virus is a potential antiviral target
    Karen Y Twu
    Institute for Cellular and Molecular Biology, Section of Molecular Genetics and Microbiology, University of Texas at Austin, 2500 Speedway, Austin, Texas 78712, USA
    J Virol 80:3957-65. 2006
    ....
  20. pmc 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
    Ji Young Min
    Section of Molecular Genetics and Microbiology, Institute for Cellular and Molecular Biology, University of Texas, Austin, TX 78712, USA
    Proc Natl Acad Sci U S A 103:7100-5. 2006
    ....
  21. ncbi Retinoic acid inducible gene-I and mda-5 are involved in influenza A virus-induced expression of antiviral cytokines
    Jukka Sirén
    Department of Viral Diseases and Immunology, National Public Health Institute, Helsinki, Finland
    Microbes Infect 8:2013-20. 2006
    ..In conclusion, our results show that in epithelial cells influenza A virus-induced antiviral cytokine gene expression is triggered by RIG-I and mda-5, whose expression is positively regulated by IFN-alpha...
  22. ncbi A site on the influenza A virus NS1 protein mediates both inhibition of PKR activation and temporal regulation of viral RNA synthesis
    Ji Young Min
    Institute for Cellular and Molecular Biology, Section of Molecular Genetics and Microbiology, room 2 122, 2500 Speedway, University of Texas at Austin, Austin, TX 78712, USA
    Virology 363:236-43. 2007
    ..This interaction would occur in the nucleus, whereas PKR would bind to NS1A proteins in the cytoplasm prior to their import into the nucleus...
  23. pmc Nuclear and nucleolar targeting of influenza A virus NS1 protein: striking differences between different virus subtypes
    Krister Melen
    Department of Viral Diseases and Immunology, National Public Health Institute, Mannerheimintie 166, FIN 00300, Helsinki, Finland
    J Virol 81:5995-6006. 2007
    ..It is likely that the evolutionarily well-conserved nucleolar targeting function of NS1A protein plays a role in the pathogenesis of influenza A virus...
  24. ncbi Conserved surface features form the double-stranded RNA binding site of non-structural protein 1 (NS1) from influenza A and B viruses
    Cuifeng Yin
    Center for Advanced Biotechnology and Medicine, Northeast Structural Genomics Consortium, Department of Molecular Biology and Biochemistry, Robert Wood Johnson Medical School, Rutgers University, Piscataway, NJ 08854, USA
    J Biol Chem 282:20584-92. 2007
    ..This pocket provides a target on the surface of the NS1 protein that is potentially suitable for the development of antiviral drugs targeting both influenza A and B viruses...
  25. pmc The H5N1 influenza virus NS genes selected after 1998 enhance virus replication in mammalian cells
    Karen Y Twu
    Institute for Cellular and Molecular Biology, University of Texas at Austin, 2500 Speedway, Austin, TX 78712, USA
    J Virol 81:8112-21. 2007
    ..Consequently, these internal HK97 proteins largely compensate for the absence of F103 and M106, presumably by stabilizing the NS1A-CPSF30 complex...
  26. pmc Structural basis for suppression of a host antiviral response by influenza A virus
    Kalyan Das
    Center for Advanced Biotechnology and Medicine, Rutgers University, Piscataway, NJ 08854, USA
    Proc Natl Acad Sci U S A 105:13093-8. 2008
    ....
  27. pmc Human ISG15 conjugation targets both IFN-induced and constitutively expressed proteins functioning in diverse cellular pathways
    Chen Zhao
    Institute for Cellular and Molecular Biology, University of Texas, 2500 Speedway, Austin, TX 78712, USA
    Proc Natl Acad Sci U S A 102:10200-5. 2005
    ..By targeting a wide array of constitutively expressed proteins, ISG15 conjugation greatly extends the repertoire of cellular functions that are affected by IFN-alpha/beta...