Adaptive Mutations Resulting in Enhanced Polymerase Activity Contribute to High Virulence of Influenza A Virus in Mice

被引:80
作者
Rolling, Thierry [1 ]
Koerner, Iris [1 ]
Zimmermann, Petra [1 ]
Holz, Kristian [1 ]
Haller, Otto [1 ]
Staeheli, Peter [1 ]
Kochs, Georg [1 ]
机构
[1] Univ Freiburg, Inst Med Mikrobiol & Hyg, Dept Virol, D-79104 Freiburg, Germany
关键词
SINGLE AMINO-ACID; RNA-POLYMERASE; MOLECULAR-BASIS; CAP-BINDING; HOST-RANGE; NUCLEAR-LOCALIZATION; VIRAL POLYMERASE; PB1; PROTEIN; SUBUNIT PB2; NS1;
D O I
10.1128/JVI.00212-09
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
High virulence of influenza virus A/Puerto Rico/8/34 in mice carrying the Mx1 resistance gene was recently shown to be determined by the viral surface proteins and the viral polymerase. Here, we demonstrated high-level polymerase activity in mammalian host cells but not avian host cells and investigated which mutations in the polymerase subunits PB1, PB2, and PA are critical for increased polymerase activity and high virus virulence. Mutational analyses demonstrated that an isoleucine-to-valine change at position 504 in PB2 was the most critical and strongly enhanced the activity of the reconstituted polymerase complex. An isoleucine-to-leucine change at position 550 in PA further contributed to increased polymerase activity and high virulence, whereas all other mutations in PB1, PB2, and PA were irrelevant. To determine whether this pattern of acquired mutations represents a preferred viral strategy to gain virulence, two independent new virus adaptation experiments were performed. Surprisingly, the conservative I504V change in PB2 evolved again and was the only mutation present in an aggressive virus variant selected during the first adaptation experiment. In contrast, the virulent virus selected in the second adaptation experiment had a lysine-to-arginine change at position 208 in PB1 and a glutamate-to-glycine change at position 349 in PA. These results demonstrate that a variety of minor amino acid changes in the viral polymerase can contribute to enhanced virulence of influenza A virus. Interestingly, all virulence-enhancing mutations that we identified in this study resulted in substantially increased viral polymerase activity.
引用
收藏
页码:6673 / 6680
页数:8
相关论文
共 57 条
  • [1] Influenza virus polymerase basic protein 1 interacts with influenza virus polymerase basic protein 2 at multiple sites
    Biswas, SK
    Nayak, DP
    [J]. JOURNAL OF VIROLOGY, 1996, 70 (10) : 6716 - 6722
  • [2] Pattern of mutation in the genome of influenza A virus on adaptation to increased virulence in the mouse lung: Identification of functional themes
    Brown, EG
    Liu, H
    Kit, LC
    Baird, S
    Nesrallah, M
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (12) : 6883 - 6888
  • [3] A single mutation in the PB1-F2 of H5N1 (HK/97) and 1918 influenza A viruses contributes to increased virulence
    Conenello, Gina M.
    Zamarin, Dmitriy
    Perrone, Lucy A.
    Tumpey, Terrence
    Palese, Peter
    [J]. PLOS PATHOGENS, 2007, 3 (10) : 1414 - 1421
  • [4] Role of ran binding protein 5 in nuclear import and assembly of the influenza virus RNA polymerase complex
    Deng, Tao
    Engelhardt, Othmar G.
    Thomas, Benjamin
    Akoulitchev, Alexandre V.
    Brownlee, George G.
    Fodor, Ervin
    [J]. JOURNAL OF VIROLOGY, 2006, 80 (24) : 11911 - 11919
  • [5] Influenza A virus strains differ in sensitivity to the antiviral action of Mx-GTPase
    Dittmann, Jan
    Stertz, Silke
    Grimm, Daniel
    Steel, John
    Garcia-Sastre, Adolfo
    Haller, Otto
    Kochs, Georg
    [J]. JOURNAL OF VIROLOGY, 2008, 82 (07) : 3624 - 3631
  • [6] Association of the influenza a virus RNA-dependent RNA polymerase with cellular RNA polymerase II
    Engelhardt, OG
    Smith, M
    Fodor, E
    [J]. JOURNAL OF VIROLOGY, 2005, 79 (09) : 5812 - 5818
  • [7] Two aromatic residues in the PB2 subunit of influenza A RNA polymerase are crucial for cap binding
    Fechter, P
    Mingay, L
    Sharps, J
    Chambers, A
    Fodor, E
    Brownlee, GG
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (22) : 20381 - 20388
  • [8] A single amino acid mutation in the PA subunit of the influenza virus RNA polymerase inhibits endonucleolytic cleavage of capped RNAs
    Fodor, E
    Crow, M
    Mingay, LJ
    Deng, T
    Sharps, J
    Fechter, P
    Brownlee, GG
    [J]. JOURNAL OF VIROLOGY, 2002, 76 (18) : 8989 - 9001
  • [9] The viral polymerase mediates adaptation of an avian influenza virus to a mammalian host
    Gabriel, G
    Dauber, B
    Wolff, T
    Planz, O
    Klenk, HD
    Stech, J
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2005, 102 (51) : 18590 - 18595
  • [10] Interaction of polymerase subunit PB2 and NP with importin α1 is a determinant of host range of influenza a virus
    Gabriel, Guelsah
    Herwig, Astrid
    Klenk, Hans-Dieter
    [J]. PLOS PATHOGENS, 2008, 4 (02)