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PMID: 8666660 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The ion channel activity of the influenza virus M2 protein affects transport through the Golgi apparatus.

The Journal of cell biology ·Vol. 133 ·No. 4 ·1996-05-00 ·Pages 733-47

Sakaguchi T, Leser GP, Lamb RA

Abstract

High level expression of the M2 ion channel protein of influenza virus inhibits the rate of intracellular transport of the influenza virus hemagglutinin (HA) and that of other integral membrane glycoproteins. HA coexpressed with M2 is properly folded, is not associated with GRP78-BiP, and trimerizes with the same kinetics as when HA is expressed alone. Analysis of the rate of transport of HA from the ER to the cis and medial golgi compartments and the TGN indicated that transport through the Golgi apparatus is delayed. Uncleaved HA0 was not expressed at the cell surface, and accumulation HA at the plasma membrane was reduced to 75-80% of control cells. The delay in intracellular transport of HA on coexpression of M2 was not observed in the presence of the M2-specific ion channel blocker, amantadine, indicating that the Golgi transport delay is due to the M2 protein ion channel activity equilibrating pH between the Golgi lumen and the cytoplasm, and not due to saturation of the intracellular transport machinery. The Na+/H+ ionophore, monensin, which also equilibrates pH between the Golgi lumen and the cytoplasm, caused a similar inhibition of intracellular transport as M2 protein expression did for HA and other integral membrane glycoproteins. EM data showed a dilation of Golgi cisternae in cells expressing the M2 ion channel protein. Taken together, the data suggest a similarity of effects of M2 ion channel activity and monensin on intracellular transport through the Golgi apparatus.

MeSH Terms
Animals Biological Transport/drug effects Cattle Cell Line Cell Membrane/metabolism Chlorocebus aethiops Endoplasmic Reticulum/metabolism Endoplasmic Reticulum Chaperone BiP Golgi Apparatus/metabolism,ultrastructure HeLa Cells Hemagglutinin Glycoproteins, Influenza Virus Hemagglutinins, Viral/genetics,metabolism Hexosaminidases Humans Hydrogen-Ion Concentration Influenza A virus/metabolism Ion Channels/genetics,metabolism Kinetics Monensin/pharmacology Protein Folding Protein Precursors/metabolism Viral Matrix Proteins/genetics,metabolism
Chemicals
Endoplasmic Reticulum Chaperone BiP HSPA5 protein, human Hemagglutinin Glycoproteins, Influenza Virus Hemagglutinins, Viral Ion Channels M-protein, influenza virus M2 protein, Influenza A virus Protein Precursors Viral Matrix Proteins Monensin Hexosaminidases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Sakaguchi T
Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, Evanston, Illinois 60208-3500, USA.
Leser G P
Lamb R A
References (86)
86 references, click to expand
  1. Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase.
    Proc Natl Acad Sci U S A. 1986 Nov;83(21):8122-6 PMID: 3095828
  2. Ability of the hydrophobic fusion-related external domain of a paramyxovirus F protein to act as a membrane anchor.
    Cell. 1987 Feb 13;48(3):441-52 PMID: 3026647
  3. Semi-intact cells permeable to macromolecules: use in reconstitution of protein transport from the endoplasmic reticulum to the Golgi complex.
    Cell. 1987 Aug 14;50(4):523-34 PMID: 3038335
  4. The structure and function of the hemagglutinin membrane glycoprotein of influenza virus.
    Annu Rev Biochem. 1987;56:365-94 PMID: 3304138
  5. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.
    Science. 1988 Jan 29;239(4839):487-91 PMID: 2448875
  6. A view of acidic intracellular compartments.
    J Cell Biol. 1988 Mar;106(3):539-43 PMID: 3279044
  7. Folding, trimerization, and transport are sequential events in the biogenesis of influenza virus hemagglutinin.
    Cell. 1988 Apr 22;53(2):197-209 PMID: 3359486
  8. Polylactosaminoglycan modification of a small integral membrane glycoprotein, influenza B virus NB.
    Mol Cell Biol. 1988 Mar;8(3):1186-96 PMID: 3367907
  9. Integration of a small integral membrane protein, M2, of influenza virus into the endoplasmic reticulum: analysis of the internal signal-anchor domain of a protein with an ectoplasmic NH2 terminus.
    J Cell Biol. 1988 May;106(5):1489-98 PMID: 2836432
  10. Influenza A virus M2 protein: monoclonal antibody restriction of virus growth and detection of M2 in virions.
    J Virol. 1988 Aug;62(8):2762-72 PMID: 2455818
  11. Intracellular maturation and transport of the SV5 type II glycoprotein hemagglutinin-neuraminidase: specific and transient association with GRP78-BiP in the endoplasmic reticulum and extensive internalization from the cell surface.
    J Cell Biol. 1989 Dec;109(6 Pt 2):3273-89 PMID: 2557352
  12. Protein folding and intracellular transport: evaluation of conformational changes in nascent exocytotic proteins.
    Methods Cell Biol. 1989;32:185-206 PMID: 2691850
  13. Selective modulation of the endocytic uptake of ricin and fluid phase markers without alteration in transferrin endocytosis.
    J Biol Chem. 1990 Apr 15;265(11):6382-8 PMID: 1969411
  14. Heavy chain binding protein recognizes incompletely disulfide-bonded forms of vesicular stomatitis virus G protein.
    J Biol Chem. 1990 Apr 25;265(12):6879-83 PMID: 2157712
  15. Quality control in the endoplasmic reticulum: folding and misfolding of vesicular stomatitis virus G protein in cells and in vitro.
    J Cell Biol. 1990 Sep;111(3):857-66 PMID: 1697299
  16. Identification of a consensus motif for retention of transmembrane proteins in the endoplasmic reticulum.
    EMBO J. 1990 Oct;9(10):3153-62 PMID: 2120038
  17. Site-directed mutagenesis of virtually any plasmid by eliminating a unique site.
    Anal Biochem. 1992 Jan;200(1):81-8 PMID: 1595905
  18. Biological activity of paramyxovirus fusion proteins: factors influencing formation of syncytia.
    J Virol. 1992 Jul;66(7):4564-9 PMID: 1602561
  19. Regulation of pH by the M2 protein of influenza A viruses.
    Virus Res. 1992 Mar;22(3):247-58 PMID: 1626420
  20. Influenza virus hemagglutinin with multibasic cleavage site is activated by furin, a subtilisin-like endoprotease.
    EMBO J. 1992 Jul;11(7):2407-14 PMID: 1628614
  21. Maturation of influenza A virus hemagglutinin--estimates of the pH encountered during transport and its regulation by the M2 protein.
    Virology. 1992 Sep;190(1):11-8 PMID: 1529523
  22. The transmembrane domain of influenza A M2 protein forms amantadine-sensitive proton channels in planar lipid bilayers.
    Virology. 1992 Sep;190(1):485-9 PMID: 1382343
  23. A new method of preparing gold probes for multiple-labeling cytochemistry.
    Eur J Cell Biol. 1985 Jul;38(1):87-93 PMID: 4029177
  24. Characterization of the influenza virus M2 integral membrane protein and expression at the infected-cell surface from cloned cDNA.
    J Virol. 1985 Nov;56(2):502-11 PMID: 3840537
  25. Expression at the cell surface of biologically active fusion and hemagglutinin/neuraminidase proteins of the paramyxovirus simian virus 5 from cloned cDNA.
    Proc Natl Acad Sci U S A. 1985 Nov;82(22):7520-4 PMID: 3865176
  26. Specific binding sites for albumin restricted to plasmalemmal vesicles of continuous capillary endothelium: receptor-mediated transcytosis.
    J Cell Biol. 1986 Apr;102(4):1304-11 PMID: 3007533
  27. Posttranslational association of immunoglobulin heavy chain binding protein with nascent heavy chains in nonsecreting and secreting hybridomas.
    J Cell Biol. 1986 May;102(5):1558-66 PMID: 3084497
  28. Expression of wild-type and mutant forms of influenza hemagglutinin: the role of folding in intracellular transport.
    Cell. 1986 Sep 12;46(6):939-50 PMID: 3757030
  29. The T4 gene encodes the AIDS virus receptor and is expressed in the immune system and the brain.
    Cell. 1986 Nov 7;47(3):333-48 PMID: 3094962
  30. Influence of amantadine resistance mutations on the pH regulatory function of the M2 protein of influenza A viruses.
    Virology. 1992 Dec;191(2):541-9 PMID: 1448912
  31. Folding and assembly of viral membrane proteins.
    Virology. 1993 Apr;193(2):545-62 PMID: 8460475
  32. Retrieval of transmembrane proteins to the endoplasmic reticulum.
    J Cell Biol. 1993 Apr;121(2):317-33 PMID: 8468349
  33. Membrane glycoprotein folding, oligomerization and intracellular transport: effects of dithiothreitol in living cells.
    EMBO J. 1993 May;12(5):2151-7 PMID: 8491203
  34. Production of the M2 protein of influenza A virus in insect cells is enhanced in the presence of amantadine.
    J Gen Virol. 1993 Aug;74 ( Pt 8):1673-7 PMID: 8345358
  35. Ion channel activity of influenza A virus M2 protein: characterization of the amantadine block.
    J Virol. 1993 Sep;67(9):5585-94 PMID: 7688826
  36. Post-translational folding of influenza hemagglutinin in isolated endoplasmic reticulum-derived microsomes.
    J Biol Chem. 1993 Sep 15;268(26):19618-25 PMID: 8366105
  37. Influenza virus M2 protein ion channel activity stabilizes the native form of fowl plague virus hemagglutinin during intracellular transport.
    J Virol. 1994 Feb;68(2):911-9 PMID: 7507186
  38. Rescue of vector-expressed fowl plague virus hemagglutinin in biologically active form by acidotropic agents and coexpressed M2 protein.
    J Virol. 1994 Feb;68(2):920-6 PMID: 8289394
  39. Intracellular trafficking and activation of the furin proprotein convertase: localization to the TGN and recycling from the cell surface.
    EMBO J. 1994 Jan 1;13(1):18-33 PMID: 7508380
  40. Influenza A virus M2 ion channel protein: a structure-function analysis.
    J Virol. 1994 Mar;68(3):1551-63 PMID: 7508997
  41. Studies with cross-linking reagents on the oligomeric form of the paramyxovirus fusion protein.
    Virology. 1994 Feb 15;199(1):160-8 PMID: 8116239
  42. Structure of influenza haemagglutinin at the pH of membrane fusion.
    Nature. 1994 Sep 1;371(6492):37-43 PMID: 8072525
  43. The influenza virus hemagglutinin cytoplasmic tail is not essential for virus assembly or infectivity.
    EMBO J. 1994 Nov 15;13(22):5504-15 PMID: 7957116
  44. Direct measurement of the influenza A virus M2 protein ion channel activity in mammalian cells.
    Virology. 1994 Nov 15;205(1):133-40 PMID: 7526533
  45. Functional reconstitution in lipid vesicles of influenza virus M2 protein expressed by baculovirus: evidence for proton transfer activity.
    J Gen Virol. 1994 Dec;75 ( Pt 12):3477-84 PMID: 7527837
  46. Reconstitution of the influenza virus M2 ion channel in lipid bilayers.
    J Membr Biol. 1994 Oct;142(1):117-26 PMID: 7535851
  47. Individual roles of N-linked oligosaccharide chains in intracellular transport of the paramyxovirus SV5 fusion protein.
    Virology. 1995 May 10;209(1):250-6 PMID: 7747477
  48. The choice of alternative 5' splice sites in influenza virus M1 mRNA is regulated by the viral polymerase complex.
    Proc Natl Acad Sci U S A. 1995 Jul 3;92(14):6324-8 PMID: 7541537
  49. Abortive infection of Vero cells by an influenza A virus (FPV).
    Virology. 1995 Sep 10;212(1):225-31 PMID: 7676635
  50. Activation of the M2 ion channel of influenza virus: a role for the transmembrane domain histidine residue.
    Biophys J. 1995 Oct;69(4):1363-71 PMID: 8534806
  51. Viral and cellular small integral membrane proteins can modify ion channels endogenous to Xenopus oocytes.
    Biophys J. 1995 Nov;69(5):1819-29 PMID: 8580325
  52. Specific structural alteration of the influenza haemagglutinin by amantadine.
    EMBO J. 1990 Nov;9(11):3469-76 PMID: 2209554
  53. Product review. New mammalian expression vectors.
    Nature. 1990 Nov 1;348(6296):91-2 PMID: 2234068
  54. Structural characteristics of the M2 protein of influenza A viruses: evidence that it forms a tetrameric channel.
    Virology. 1991 Feb;180(2):617-24 PMID: 1989386
  55. Regulated M1 mRNA splicing in influenza virus-infected cells.
    J Gen Virol. 1991 Jun;72 ( Pt 6):1301-8 PMID: 1710647
  56. Influenza virus M2 integral membrane protein is a homotetramer stabilized by formation of disulfide bonds.
    Virology. 1991 Jul;183(1):32-43 PMID: 2053285
  57. A new cationic liposome reagent mediating nearly quantitative transfection of animal cells.
    Biotechniques. 1991 Apr;10(4):520-5 PMID: 1867862
  58. Alterations to influenza virus hemagglutinin cytoplasmic tail modulate virus infectivity.
    J Virol. 1992 Feb;66(2):790-803 PMID: 1309913
  59. Studies on the fusion peptide of a paramyxovirus fusion glycoprotein: roles of conserved residues in cell fusion.
    J Virol. 1992 Apr;66(4):2443-55 PMID: 1548771
  60. 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. 1992 May;188(1):14-24 PMID: 1566569
  61. Influenza virus M2 protein has ion channel activity.
    Cell. 1992 May 1;69(3):517-28 PMID: 1374685
  62. Unpacking the incoming influenza virus.
    Cell. 1992 May 15;69(4):577-8 PMID: 1375129
  63. Deficient uridine diphosphate-N-acetylglucosamine:glycoprotein N-acetylglucosaminyltransferase activity in a clone of Chinese hamster ovary cells with altered surface glycoproteins.
    J Biol Chem. 1975 May 10;250(9):3303-9 PMID: 1168193
  64. The synthesis of sendai virus polypeptides in infected cells.
    Virology. 1976 Jan;69(1):116-31 PMID: 174287
  65. Evidence from studies with a cross-linking reagent that the haemagglutinin of influenza virus is a trimer.
    Virology. 1977 Jun 15;79(2):446-8 PMID: 867833
  66. Monovalent ionophores inhibit secretion of procollagen and fibronectin from cultured human fibroblasts.
    Proc Natl Acad Sci U S A. 1979 Apr;76(4):1868-72 PMID: 377287
  67. Segment 8 of the influenza virus genome is unique in coding for two polypeptides.
    Proc Natl Acad Sci U S A. 1979 Oct;76(10):4908-12 PMID: 291907
  68. Intracellular transport of secretory and membrane proteins in hepatoma cells infected by vesicular stomatitis virus.
    Cell. 1980 Dec;22(3):709-17 PMID: 6257395
  69. Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3 A resolution.
    Nature. 1981 Jan 29;289(5796):366-73 PMID: 7464906
  70. Differential effect of monensin on enveloped viruses that form at distinct plasma membrane domains.
    J Cell Biol. 1981 Jun;89(3):700-5 PMID: 6265471
  71. Identification of a second protein (M2) encoded by RNA segment 7 of influenza virus.
    Virology. 1981 Jul 30;112(2):729-37 PMID: 7257188
  72. Changes in the conformation of influenza virus hemagglutinin at the pH optimum of virus-mediated membrane fusion.
    Proc Natl Acad Sci U S A. 1982 Feb;79(4):968-72 PMID: 6951181
  73. Effects of monensin on morphogenesis and infectivity of Friend murine leukemia virus.
    J Virol. 1982 Jun;42(3):1067-75 PMID: 7097856
  74. Spliced and unspliced messenger RNAs synthesized from cloned influenza virus M DNA in an SV40 vector: expression of the influenza virus membrane protein (M1).
    Virology. 1982 Dec;123(2):237-56 PMID: 6184879
  75. Dissection of the Golgi complex. I. Monensin inhibits the transport of viral membrane proteins from medial to trans Golgi cisternae in baby hamster kidney cells infected with Semliki Forest virus.
    J Cell Biol. 1983 Mar;96(3):835-50 PMID: 6682112
  76. Perturbation of vesicular traffic with the carboxylic ionophore monensin.
    Cell. 1983 Apr;32(4):1026-8 PMID: 6340834
  77. Modulation of glycosylation and transport of viral membrane glycoproteins by a sodium ionophore.
    J Cell Biol. 1983 Sep;97(3):659-68 PMID: 6309867
  78. Intracellular transport of influenza virus hemagglutinin to the apical surface of Madin-Darby canine kidney cells.
    J Cell Biol. 1984 Jan;98(1):308-19 PMID: 6707094
  79. Structure of branched lactosaminoglycan, the carbohydrate moiety of band 3 isolated from adult human erythrocytes.
    J Biol Chem. 1984 Jul 10;259(13):8260-73 PMID: 6736034
  80. New evidence of the substrate specificity of endo-beta-N-acetylglucosaminidase D.
    J Biochem. 1984 Apr;95(4):1209-13 PMID: 6430882
  81. Visualization of acidic organelles in intact cells by electron microscopy.
    Proc Natl Acad Sci U S A. 1984 Aug;81(15):4838-42 PMID: 6146980
  82. A clathrin-coated, Golgi-related compartment of the insulin secreting cell accumulates proinsulin in the presence of monensin.
    Cell. 1984 Nov;39(1):39-47 PMID: 6386182
  83. Fusion protein of the paramyxovirus simian virus 5: nucleotide sequence of mRNA predicts a highly hydrophobic glycoprotein.
    Proc Natl Acad Sci U S A. 1984 Nov;81(21):6706-10 PMID: 6093114
  84. Influenza virus M2 protein is an integral membrane protein expressed on the infected-cell surface.
    Cell. 1985 Mar;40(3):627-33 PMID: 3882238
  85. Cell surface glycoconjugates as onco-differentiation markers in hematopoietic cells.
    Biochim Biophys Acta. 1985;780(2):119-50 PMID: 3890942
  86. ATP-coupled transport of vesicular stomatitis virus G protein between the endoplasmic reticulum and the Golgi.
    J Biol Chem. 1986 Nov 5;261(31):14681-9 PMID: 3021750
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.133.4.733
Published
1996-05-00
Pages
733-47
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120830
Subset
IM
Grants
NIAID NIH HHS · AI-20201 · United States
External Links
PubMed source DOI: 10.1083/jcb.133.4.733 PMC Full Text View journal details
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