Archives of Virology

, 154:147 | Cite as

Stability and function of the influenza A virus M2 ion channel protein is determined by both extracellular and cytoplasmic domains

Brief Report


A series of M2/NB chimeras were used to investigate the ion channel activity of the IAV M2 protein. Replacing the M2 cytoplasmic domain with the equivalent NB domain (AAB chimera) did not influence ion channel activity, while replacement of N-terminal domains (BAA and BAB chimeras) resulted in loss of activity. Extension of the M2 protein N-terminal domain resulted in full restoration of ion channel activity in BAA chimeras but only partial restoration in BAB. While not directly involved in ion channel activity, the N- and C-terminals of M2 are important for stabilization of the transmembrane domain structure.


Influenza Cytoplasmic Tail Rimantadine Internal Viral Protein Vaccinia Virus vTF7 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



trans Golgi network




Influenza A virus


Influenza B virus



We thank Michael S. Bennet and Seti Grambas for excellent assistance. This research was partially supported by the Slovak Research and Development Agency (grant no. APVV-51-004105) and the VEGA-Grant Agency of Science (grant no. 2/6152/06).


  1. 1.
    Lamb RA, Zebedee SL, Richardson CD (1985) Influenza virus M2 protein is an integral membrane protein expressed on the infected-cell surface. Cell 40:627–633PubMedCrossRefGoogle Scholar
  2. 2.
    Betakova T, Nermut MV, Hay AJ (1996) The NB protein is an integral component of the membrane of influenza B virus. J Gen Virol 77:2689–2694PubMedCrossRefGoogle Scholar
  3. 3.
    von Heijne G (1988) Transcending the impenetrable: how proteins come to terms with membranes. Biochim Biophys Acta 947:307–333Google Scholar
  4. 4.
    Zebedee SL, Lamb RA (1988) Influenza A virus M2 protein: monoclonal antibody restriction of growth and detection of M2 in virions. J Virol 62:2762–2772PubMedGoogle Scholar
  5. 5.
    Sugrue RJ, Belshe RB, Hay AJ (1990) Palmitoylation of the influenza A virus M2 protein. Virology 179:51–56PubMedCrossRefGoogle Scholar
  6. 6.
    Sugrue RJ, Hay AJ (1991) Structural characteristics of the M2 protein of influenza A viruses: evidence that it forms a tetrameric channel. Virology 180:617–624PubMedCrossRefGoogle Scholar
  7. 7.
    Holsinger LJ, Lamb RA (1991) Influenza virus M2 integral membrane protein is a homotetramer stabilized by formation of disulfide bonds. Virology 183:32–43PubMedCrossRefGoogle Scholar
  8. 8.
    Martin K, Helenius A (1991) Nuclear transport of influenza virus ribonucleoproteins: the viral matrix protein (M1) promotes export and inhibits import. Cell 67:117–130PubMedCrossRefGoogle Scholar
  9. 9.
    Helenius A (1992) Unpacking the incoming influenza virus. Cell 69:577–578PubMedCrossRefGoogle Scholar
  10. 10.
    Ciampor F, Bayley PM, Nermut MV, Hirst EM, Sugrue RJ, Hay AJ (1992) Evidence that the amantadine-induced M2-mediated conversion of influenza A virus hemagglutinin to the low pH conformation occurs in an acidic trans-Golgi compartment. Virology 188:14–24PubMedCrossRefGoogle Scholar
  11. 11.
    Grambas S, Bennet MS, Hay AJ (1992) Influence of amantadine resistance mutations on the pH regulatory function of the M2 protein of influenza A virus. Virology 191:541–549PubMedCrossRefGoogle Scholar
  12. 12.
    Grambas S, Hay AJ (1992) Maturation of influenza A virus hemagglutinin—estimates of the pH encountered during transport and its regulation by the M2 protein. Virology 190:11–18PubMedCrossRefGoogle Scholar
  13. 13.
    Chizhmakov IV, Geraghty FM, Ogden DC, Hayhurst A, Antoniou M, Hay AJ (1996) Selective proton permeability and pH regulation of the influenza virus M2 channel expressed in mouse erythroleukemia cells. J Physiol 494:329–336PubMedGoogle Scholar
  14. 14.
    Mould JA, Drury JE, Frings SM, Kaupp UB, Pekosz A, Lamb RA, Pinto LH (2000) Permeation and activation of the M2 ion channel of influenza A virus. J Biol Chem 275:31038–31050PubMedCrossRefGoogle Scholar
  15. 15.
    Betakova T, Kollerova E (2006) pH modulating activity of ion channels of influenza A, B, and C viruses. Acta Virol 50:187–193PubMedGoogle Scholar
  16. 16.
    Betakova T, Hay AJ (2007) Evidence that the CM2 protein of influenza C virus can modify the pH of the exocytic pathway of transfected cells. J Gen Virol 88:2291–2296PubMedCrossRefGoogle Scholar
  17. 17.
    Hatta M, Kawaoka Y (2003) The NB protein of influenza B virus is not necessary for virus replication in vitro. J Virol 77:6050–6054PubMedCrossRefGoogle Scholar
  18. 18.
    Williams MA, Lamb RA (1986) Determination of the orientation of an integral membrane protein and sites of glycosylation by oligonucleotide-direct mutagenesis: influenza B virus NB glycoprotein lacks a cleavable signal sequence and has an extracellular NH2 -terminal region. Mol Cell Biol 6:4317–4328PubMedGoogle Scholar
  19. 19.
    Williams MA, Lamb RA (1988) Polylactosaminoglycan modification of small integral membrane glycoprotein, influenza B virus NB. Mol Cell Biol 8:1186–1196PubMedGoogle Scholar
  20. 20.
    Tobler K, Kelly ML, Pinto LH, Lamb RA (1999) Effect of cytoplasmic tail truncations on the activity of the ion channel of influenza A virus. J Virol 73:9695–9701PubMedGoogle Scholar
  21. 21.
    McCown MF, Pekosz A (2005) The influenza A virus M2 cytoplasmic tail is required for infectious virus production and efficient genome packaging. J Virol 79:3595–3605PubMedCrossRefGoogle Scholar
  22. 22.
    McCown MF, Pekosz A (2006) Distinct domains of the influenza A virus M2 protein cytoplasmic tail mediate binding to the M1 protein and facilitate infectious virus production. J Virol 80:8178–8189PubMedCrossRefGoogle Scholar
  23. 23.
    Chen BJ, Leser GP, Jackson D, Lamb RA (2008) The influenza virus M2 protein cytoplasmic tail interacts with M1 protein and influences virus assembly at the site of virus budding. J Virol 82:10059–10070PubMedCrossRefGoogle Scholar
  24. 24.
    Iwatsuki-Horimoto K, Horimoto T, Noda T, Kiso M, Maeda J, Watanabe S, Muramoto Y, Fujii K, Kawaoka Y (2006) The cytoplasmic tail of the influenza A virus M2 protein plays a role in viral assembly. J Virol 80:5233–5240PubMedCrossRefGoogle Scholar
  25. 25.
    Park EK, Castrucci MR, Portner A, Kawaoka Y (1998) The M2 ectodomain is important for its incorporation into influenza A virions. J Virol 72:2449–2455PubMedGoogle Scholar
  26. 26.
    Liu W, Li H, Chen YH (2003) N-terminus of M2 protein could induce antibodies with inhibitory activity against influenza virus replication. FEMS Immunol Med Microbiol 35:141–146PubMedCrossRefGoogle Scholar
  27. 27.
    Betakova T, Ciampor F, Hay A (2005) Influence of residue 44 on the activity of the M2 proton channel of influenza A virus. J Gen Virol 86:181–184PubMedCrossRefGoogle Scholar
  28. 28.
    Fuerst TR, Niles EG, Studier FW, Moss B (1986) Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase. Proc Natl Acad Sci USA 83:8122–8126PubMedCrossRefGoogle Scholar
  29. 29.
    Forrest LR, Tieleman DP, Sansom MP (1999) Defining the transmembrane helix of M2 protein from influenza A by molecular dynamics simulations in a lipid bilayer. Biophys J 76:1886–1896PubMedCrossRefGoogle Scholar
  30. 30.
    Forrest LR, Kukol A, Arkin IT, Tielemen DP, Sansom MP (2000) Exploring models of the influenza A M2 channel: MD simulation in a phospholipid bilayer. Biophys J 78:55–69PubMedGoogle Scholar
  31. 31.
    Kollerova E, Betakova T (2007) Influence of extracellular and cytoplasmic domains of M2 proton channel of influenza A virus on its activity. Acta Virol 51:73–78Google Scholar
  32. 32.
    Schnell JR, Chou JJ (2008) Structure and mechanism of the M2 proton channel of influenza A virus. Nature 451:591–595PubMedCrossRefGoogle Scholar
  33. 33.
    Nguyen PA, Soto CS, Polishchuk A, Caputo GA, Tatko CD, Ma C, Ohigashi Y, Pinto LH, DeGrado WF, Howard KP (2008) pH-Induced conformational change of the influenza M2 protein C-terminal domain. Biochem 47:9934–9936CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.Institute of VirologySlovak Academy of ScienceBratislavaSlovakia
  2. 2.National Institute for Medical ResearchLondonUK

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