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Na+/K+-ATPase β1 subunit interacts with M2 proteins of influenza A and B viruses and affects the virus replication

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Abstract

Interplay between the host and influenza virus has a pivotal role for the outcome of infection. The matrix proteins M2/BM2 from influenza (A and B) viruses are small type III integral membrane proteins with a single transmembrane domain, a short amino-terminal ectodomain and a long carboxy-terminal cytoplasmic domain. They function as proton channels, mainly forming a membrane-spanning pore through the transmembrane domain tetramer, and are essential for virus assembly and release of the viral genetic materials in the endosomal fusion process. However, little is known about the host factors which interact with M2/BM2 proteins and the functions of the long cytoplasmic domain are currently unknown. Starting with yeast two-hybrid screening and applying a series of experiments we identified that the β1 subunit of the host Na+/K+-ATPase β1 subunit (ATP1B1) interacts with the cytoplasmic domain of both the M2 and BM2 proteins. A stable ATP1B1 knockdown MDCK cell line was established and we showed that the ATP1B1 knockdown suppressed influenza virus A/WSN/33 replication, implying that the interaction is crucial for influenza virus replication in the host cell. We propose that influenza virus M2/BM2 cytoplasmic domain has an important role in the virus-host interplay and facilitates virus replication.

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References

  1. Gao G F, Sun Y P. It is not just AIV: From avian to swine-origin influenza virus. Sci China Life Sci, 2010, 53:151–153 10.1007/s11427-010-0017-4, 20596968

    Article  PubMed  Google Scholar 

  2. Kang S, Yang I S, Lee J Y, et al. Epidemiologic study of human influenza virus infection in South Korea from 1999 to 2007: origin and evolution of A/Fujian/411/2002-like strains. J Clin Microbiol, 2010, 48:2177–2185 1:CAS:528:DC%2BC3cXpsVGntL4%3D, 10.1128/JCM.00209-10, 20392920

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  3. Khanna M, Gupta N, Gupta A, et al. Influenza A (H1N1) 2009: a pandemic alarm. J Biosci, 2009, 34:481–489 10.1007/s12038-009-0053-z, 19805908

    Article  PubMed  Google Scholar 

  4. Zhang W, Qi J X, Shi Y, et al. Crystal structure of the swine-origin A (H1N1)—2009 influenza A virus hemagglutinin (HA) reveals similar antigenicity to that of the 1918 pandemic virus. Protein & Cell, 2010, 1:459–467 1:CAS:528:DC%2BC3cXosleluro%3D, 10.1007/s13238-010-0059-1

    Article  CAS  Google Scholar 

  5. Liu D, Liu X L, Yan J H, et al. Interspecies transmission and host restriction of avian H5N1 influenza virus. Sci China Ser C-Life Sci, 2009, 52:428–438 10.1007/s11427-009-0062-z

    Article  Google Scholar 

  6. Knipe D M, Howley P M. Fields Virology. Philadelphia: Lippincott Williams & Wilkins, a Wolters Kluwer Business, 2007

    Google Scholar 

  7. Lamb R A, Lai C J, Choppin P W. Sequences of mRNAs derived from genome RNA segment 7 of influenza virus: colinear and interrupted mRNAs code for overlapping proteins. Proc Natl Acad Sci USA, 1981, 78:4170–4174 1:CAS:528:DyaL3MXlvF2hsro%3D, 10.1073/pnas.78.7.4170, 6945577

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  8. Sugrue R J, Belshe R B, Hay A J. Palmitoylation of the influenza A virus M2 protein. Virology, 1990, 179:51–56 1:CAS:528:DyaK3cXmt1Oktr0%3D, 10.1016/0042-6822(90)90272-S, 2219738

    Article  PubMed  CAS  Google Scholar 

  9. Holsinger L J, Lamb R A. Influenza virus M2 integral membrane protein is a homotetramer stabilized by formation of disulfide bonds. Virology, 1991, 183:32–43 1:CAS:528:DyaK3MXksVCmtL8%3D, 10.1016/0042-6822(91)90115-R, 2053285

    Article  PubMed  CAS  Google Scholar 

  10. Thomas J M, Stevens M P, Percy N, et al. Phosphorylation of the M2 protein of influenza A virus is not essential for virus viability. Virology, 1998, 252:54–64 1:CAS:528:DyaK1cXotFSitLY%3D, 10.1006/viro.1998.9384, 9875317

    Article  PubMed  CAS  Google Scholar 

  11. Horvath C M, Williams M A, Lamb R A. Eukaryotic coupled translation of tandem cistrons: identification of the influenza B virus BM2 polypeptide. EMBO J, 1990, 9:2639–2647 1:CAS:528:DyaK3cXlt1yhu7c%3D, 2114979

    PubMed  CAS  PubMed Central  Google Scholar 

  12. Odagiri T, Hong J, Ohara Y. The BM2 protein of influenza B virus is synthesized in the late phase of infection and incorporated into virions as a subviral component. J Gen Virol, 1999, 80:2573–2581 1:CAS:528:DyaK1MXmsVygsb4%3D, 10573149

    Article  PubMed  CAS  Google Scholar 

  13. Park E K, Castrucci M R, Portner A, et al. The M2 ectodomain is important for its incorporation into influenza A virions. J Virol, 1998, 72:2449–2455 1:CAS:528:DyaK1cXhtFegsbc%3D, 9499106

    PubMed  CAS  PubMed Central  Google Scholar 

  14. Paterson R G, Takeda M, Ohigashi Y, et al. Influenza B virus BM2 protein is an oligomeric integral membrane protein expressed at the cell surface. Virology, 2003, 306:7–17 1:CAS:528:DC%2BD3sXhs1Kntrg%3D, 10.1016/S0042-6822(02)00083-1, 12620792

    Article  PubMed  CAS  Google Scholar 

  15. Watanabe S, Imai M, Ohara Y, et al. Influenza B virus BM2 protein is transported through the trans-Golgi network as an integral membrane protein. J Virol, 2003, 77:10630–10637 1:CAS:528:DC%2BD3sXnsFymt7s%3D, 10.1128/JVI.77.19.10630-10637.2003, 12970447

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  16. Wang J, Pielak R M, McClintock M A, et al. Solution structure and functional analysis of the influenza B proton channel. Nat Struct Mol Biol, 2009, 16:1267–1271 1:CAS:528:DC%2BD1MXhtlOrurrO, 10.1038/nsmb.1707, 19898475

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  17. Sugrue R J, Hay A J. Structural characteristics of the M2 protein of influenza A viruses: evidence that it forms a tetrameric channel. Virology, 1991, 180:617–624 1:CAS:528:DyaK3MXnvVeluw%3D%3D, 10.1016/0042-6822(91)90075-M, 1989386

    Article  PubMed  CAS  Google Scholar 

  18. Pinto L H, Holsinger L J, Lamb R A. Influenza virus M2 protein has ion channel activity. Cell, 1992, 69:517–528 1:CAS:528:DyaK38Xks1Cqtrg%3D, 10.1016/0092-8674(92)90452-I, 1374685

    Article  PubMed  CAS  Google Scholar 

  19. Henkel J R, Gibson G A, Poland P A, et al. Influenza M2 proton channel activity selectively inhibits trans-Golgi network release of apical membrane and secreted proteins in polarized Madin-Darby canine kidney cells. J Cell Biol, 2000, 148:495–504 1:CAS:528:DC%2BD3cXhtFSqtrc%3D, 10.1083/jcb.148.3.495, 10662775

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  20. Takeda M, Pekosz A, Shuck K, et al. Influenza a virus M2 ion channel activity is essential for efficient replication in tissue culture. J Virol, 2002, 76:1391–1399 1:CAS:528:DC%2BD38XnvFCqsg%3D%3D, 10.1128/JVI.76.3.1391-1399.2002, 11773413

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  21. Mould J A, Paterson R G, Takeda M, et al. Influenza B virus BM2 protein has ion channel activity that conducts protons across membranes. Dev Cell, 2003, 5:175–184 1:CAS:528:DC%2BD3sXmt1Smsb0%3D, 10.1016/S1534-5807(03)00190-4, 12852861

    Article  PubMed  CAS  Google Scholar 

  22. Pinto L H, Lamb R A. The M2 proton channels of influenza A and B viruses. J Biol Chem, 2006, 281:8997–9000 1:CAS:528:DC%2BD28XjtVSktrg%3D, 10.1074/jbc.R500020200, 16407184

    Article  PubMed  CAS  Google Scholar 

  23. McCown M F, Pekosz A. The influenza A virus M2 cytoplasmic tail is required for infectious virus production and efficient genome packaging. J Virol, 2005, 79:3595–3605 1:CAS:528:DC%2BD2MXit1ehs7Y%3D, 10.1128/JVI.79.6.3595-3605.2005, 15731254

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  24. McCown M F, Pekosz A. Distinct domains of the influenza a virus M2 protein cytoplasmic tail mediate binding to the M1 protein and facilitate infectious virus production. J Virol, 2006, 80:8178–8189 1:CAS:528:DC%2BD28Xot1yrtLY%3D, 10.1128/JVI.00627-06, 16873274

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  25. Chen B J, Leser G P, Jackson D, et al. The influenza virus M2 protein cytoplasmic tail interacts with the M1 protein and influences virus assembly at the site of virus budding. J Virol, 2008, 82:10059–10070 1:CAS:528:DC%2BD1cXht1amtrvM, 10.1128/JVI.01184-08, 18701586

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  26. Hatta M, Goto H, Kawaoka Y. Influenza B virus requires BM2 pro tein for replication. J Virol, 2004, 78:5576–5583 1:CAS:528:DC%2BD2cXksFCgs7w%3D, 10.1128/JVI.78.11.5576-5583.2004, 15140954

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  27. Imai M, Watanabe S, Ninomiya A, et al. Influenza B virus BM2 protein is a crucial component for incorporation of viral ribonucleoprotein complex into virions during virus assembly. J Virol, 2004, 78:11007–11015 1:CAS:528:DC%2BD2cXotlKju70%3D, 10.1128/JVI.78.20.11007-11015.2004, 15452221

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  28. Zhou Z, Jiang X, Liu D, et al. Autophagy is involved in influenza A virus replication. Autophagy, 2009, 5:321–328 1:CAS:528:DC%2BD1MXnvVyku7Y%3D, 10.4161/auto.5.3.7406, 19066474

    Article  PubMed  CAS  Google Scholar 

  29. Rossman J S, Lamb R A. Autophagy, apoptosis, and the influenza virus M2 protein. Cell Host Microbe, 2009, 6:299–300 1:CAS:528:DC%2BD1MXhsVCjtr3J, 10.1016/j.chom.2009.09.009, 19837369

    Article  PubMed  CAS  Google Scholar 

  30. Skou J C, Esmann M. The Na,K-ATPase. J Bioenerg Biomembr, 1992, 24:249–261 1:CAS:528:DyaK38XltVKrsbg%3D, 1328174

    PubMed  CAS  Google Scholar 

  31. Mobasheri A, Avila J, Cozar-Castellano I, et al. Na+,K+-ATPase isozyme diversity; comparative biochemistry and physiological implications of novel functional interactions. Biosci Rep, 2000, 20:51–91 1:CAS:528:DC%2BD3cXmtFymu7w%3D, 10.1023/A:1005580332144, 10965965

    Article  PubMed  CAS  Google Scholar 

  32. Emanuel J R, Garetz S, Stone L, et al. Differential expression of Na+,K+-ATPase alpha- and beta-subunit mRNAs in rat tissues and cell lines. Proc Natl Acad Sci USA, 1987, 84:9030–9034 1:CAS:528:DyaL1cXht1Gls78%3D, 10.1073/pnas.84.24.9030, 2827165

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  33. Miller R P, Farley R A. All three potential N-glycosylation sites of the dog kidney (Na++K+)-ATPase beta-subunit contain oligosaccharide. Biochim Biophys Acta, 1988, 954:50–57 1:CAS:528:DyaL1cXktVejsrc%3D, 2833926

    Article  PubMed  CAS  Google Scholar 

  34. Miller R P, Farley R A. Beta subunit of (Na++K+)-ATPase contains three disulfide bonds. Biochemistry, 1990, 29:1524–1532 1:CAS:528:DyaK3cXnt1yktQ%3D%3D, 10.1021/bi00458a025, 2159340

    Article  PubMed  CAS  Google Scholar 

  35. Noguchi S, Higashi K, Kawamura M. A possible role of the betasubunit of (Na,K)-ATPase in facilitating correct assembly of the alphasubunit into the membrane. J Biol Chem, 1990, 265:15991–15995 1:CAS:528:DyaK3cXls1yjsbY%3D, 2168428

    PubMed  CAS  Google Scholar 

  36. Rajasekaran S A, Gopal J, Willis D, et al. Na,K-ATPase beta1-subunit increases the translation efficiency of the alpha1-subunit in MSV-MDCK cells. Mol Biol Cell, 2004, 15:3224–3232 1:CAS:528:DC%2BD2cXlslWnsL0%3D, 10.1091/mbc.E04-03-0222, 15133131

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  37. Shoshani L, Contreras R G, Roldan M L, et al. The polarized expression of Na+,K+-ATPase in epithelia depends on the association between beta-subunits located in neighboring cells. Mol Biol Cell, 2005, 16:1071–1081 1:CAS:528:DC%2BD2MXit1Srsbs%3D, 10.1091/mbc.E04-03-0267, 15616198

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  38. Yoshimura S H, Iwasaka S, Schwarz W, et al. Fast degradation of the auxiliary subunit of Na+/K+-ATPase in the plasma membrane of HeLa cells. J Cell Sci, 2008, 121:2159–2168 1:CAS:528:DC%2BD1cXptF2jtbw%3D, 10.1242/jcs.022905, 18522992

    Article  PubMed  CAS  Google Scholar 

  39. Kitamura N, Ikekita M, Sato T, et al. Mouse Na+/K+-ATPase beta1-subunit has a K+-dependent cell adhesion activity for beta-GlcNActerminating glycans. Proc Natl Acad Sci USA, 2005, 102:2796–2801 1:CAS:528:DC%2BD2MXitVSksL0%3D, 10.1073/pnas.0409344102, 15705719

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  40. Rajasekaran S A, Palmer L G, Quan K, et al. Na,K-ATPase beta-subunit is required for epithelial polarization, suppression of invasion, and cell motility. Mol Biol Cell, 2001, 12:279–295 1:CAS:528:DC%2BD3MXhtlGjtL0%3D, 11179415

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  41. Pratscher B, Friedrich C, Goger W, et al. Characterization of NKIP: a novel, Na+/K+-ATPase interacting protein mediates neural differentiation and apoptosis. Exp Cell Res, 2008, 314:463–477 1:CAS:528:DC%2BD1cXpsFOruw%3D%3D, 10.1016/j.yexcr.2007.11.013, 18096156

    Article  PubMed  CAS  Google Scholar 

  42. Jha S, Dryer S E. The beta1 subunit of Na+/K+-ATPase interacts with BKCa channels and affects their steady-state expression on the cell surface. FEBS Lett, 2009, 583:3109–3114 1:CAS:528:DC%2BD1MXht1aisrnP, 10.1016/j.febslet.2009.08.039, 19729011

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  43. Kirley T L. Determination of three disulfide bonds and one free sulfhydryl in the beta subunit of (Na,K)-ATPase. J Biol Chem, 1989, 264:7185–7192 1:CAS:528:DyaL1MXktVeht7g%3D, 2540179

    PubMed  CAS  Google Scholar 

  44. Gannage M, Dormann D, Albrecht R, et al. Matrix protein 2 of influenza A virus blocks autophagosome fusion with lysosomes. Cell Host Microbe, 2009, 6:367–380 1:CAS:528:DC%2BD1MXhsVCjtrrN, 10.1016/j.chom.2009.09.005, 19837376

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  45. Rossman J S, Jing X, Leser G P, et al. Influenza virus m2 ion channel protein is necessary for filamentous virion formation. J Virol, 2010, 84:5078–5088 1:CAS:528:DC%2BC3cXmtVWgsL8%3D, 10.1128/JVI.00119-10, 20219914

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  46. Wilson P D, Devuyst O, Li X, et al. Apical plasma membrane mispolarization of NaK-ATPase in polycystic kidney disease epithelia is associated with aberrant expression of the beta2 isoform. Am J Pathol, 2000, 156:253–268 1:CAS:528:DC%2BD3cXns1yksg%3D%3D, 10623674

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  47. Ichinohe T, Pang I K, Iwasaki A. Influenza virus activates inflammasomes via its intracellular M2 ion channel. Nat Immunol, 2010, 11:404–410 1:CAS:528:DC%2BC3cXksFaks7Y%3D, 10.1038/ni.1861, 20383149

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  48. Xie Z, Wang Y, Liu G, et al. Similarities and differences between the properties of native and recombinant Na+/K+-ATPases. Arch Biochem Biophys, 1996, 330:153–162 1:CAS:528:DyaK28Xjtlait7o%3D, 10.1006/abbi.1996.0237, 8651690

    Article  PubMed  CAS  Google Scholar 

  49. Noguchi S, Higashi K, Kawamura M. Assembly of the alpha-subunit of Torpedo californica Na+/K(+)-ATPase with its pre-existing beta-subunit in Xenopus oocytes. Biochim Biophys Acta, 1990, 1023:247–253 1:CAS:528:DyaK3cXitlWrtbw%3D, 10.1016/0005-2736(90)90420-S, 2158350

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China

    ShuoFu Mi, Yan Li, JingHua Yan & George Fu Gao

  2. Graduate University, Chinese Academy of Sciences, Beijing, 100049, China

    ShuoFu Mi, Yan Li & George Fu Gao

  3. Beijing Institutes of Life Sciences, Chinese Academy of Sciences, Beijing, 100101, China

    George Fu Gao

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  1. ShuoFu Mi
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Mi, S., Li, Y., Yan, J. et al. Na+/K+-ATPase β1 subunit interacts with M2 proteins of influenza A and B viruses and affects the virus replication. Sci. China Life Sci. 53, 1098–1105 (2010). https://doi.org/10.1007/s11427-010-4048-7

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