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PMID: 8858163 Published · ppublish English Journal Article

Dilation of the influenza hemagglutinin fusion pore revealed by the kinetics of individual cell-cell fusion events.

The Journal of cell biology ·Vol. 135 ·No. 1 ·1996-10-00 ·Pages 63-71

Blumenthal R, Sarkar DP, Durell S, Howard DE, Morris SJ

Abstract

We have monitored kinetics of fusion between cell pairs consisting of a single influenza hemaglutinin (HA)-expressing cell and a single erythrocyte (RBC) that had been labeled with both a fluorescent lipid (Dil) in the membrane and a fluorescent solute (calcein) in the aqueous space. Initial fusion pore opening between the RBC and HA-expressing cell produced a change in RBC membrane potential (delta psi) that was monitored by a decrease in Dil fluorescence. This event was followed by two distinct stages of fusion pore dilation: the flux of fluorescent lipid (phi L) and the flux of a large aqueous fluorescent dye (phi s). We have analyzed the kinetics of events that occur as a result of transitions between a fusion pore (FP) and a solute permissive fusion pore (FPs). Our data are consistent with a fusion pore comprising six HA trimers.

MeSH Terms
3T3 Cells Animals CHO Cells Cell Fusion Cricetinae Erythrocyte Membrane/physiology Fluoresceins Fluorescent Dyes Hemagglutinin Glycoproteins, Influenza Virus/physiology Humans Kinetics Membrane Fusion/physiology Membrane Potentials Mice Microscopy, Video Temperature
Chemicals
Fluoresceins Fluorescent Dyes Hemagglutinin Glycoproteins, Influenza Virus fluorexon
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Blumenthal R
Section on Membrane Structure and Function, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA. [email protected]
Sarkar D P
Durell S
Howard D E
Morris S J
References (45)
45 references, click to expand
  1. Role of the fusion peptide sequence in initial stages of influenza hemagglutinin-induced cell fusion.
    J Biol Chem. 1993 May 5;268(13):9267-74 PMID: 8387488
  2. The exocytotic fusion pore interface: a model of the site of neurotransmitter release.
    Mol Membr Biol. 1995 Jan-Mar;12(1):151-6 PMID: 7767376
  3. Role of target membrane structure in fusion with influenza virus: effect of modulating erythrocyte transbilayer phospholipid distribution.
    Membr Biochem. 1993 Jan-Mar;10(1):3-15 PMID: 8510561
  4. Kinetics of cell fusion mediated by viral spike glycoproteins.
    Methods Enzymol. 1993;221:42-58 PMID: 8361385
  5. Enhancement of viral fusion by nonadsorbing polymers.
    Biophys J. 1993 Jul;65(1):528-34 PMID: 7690263
  6. Lipid-anchored influenza hemagglutinin promotes hemifusion, not complete fusion.
    Cell. 1994 Jan 28;76(2):383-91 PMID: 8293471
  7. Energetics of intermediates in membrane fusion: comparison of stalk and inverted micellar intermediate mechanisms.
    Biophys J. 1993 Nov;65(5):2124-40 PMID: 8298039
  8. The synaptic vesicle cycle: a cascade of protein-protein interactions.
    Nature. 1995 Jun 22;375(6533):645-53 PMID: 7791897
  9. Membrane potential-driven translocation of a lipid-conjugated rhodamine.
    Biochim Biophys Acta. 1995 Jul 26;1237(2):121-6 PMID: 7632704
  10. Determination of membrane potentials in human and Amphiuma red blood cells by means of fluorescent probe.
    J Physiol. 1974 Jun;239(3):519-52 PMID: 4851321
  11. Membrane fusion during secretion: cortical granule exocytosis in sex urchin eggs as studied by quick-freezing and freeze-fracture.
    J Cell Biol. 1979 Oct;83(1):91-108 PMID: 574515
  12. Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3 A resolution.
    Nature. 1981 Jan 29;289(5796):366-73 PMID: 7464906
  13. Cell fusion by Semliki Forest, influenza, and vesicular stomatitis viruses.
    J Cell Biol. 1981 Jun;89(3):674-9 PMID: 6265470
  14. Capacitance measurements reveal stepwise fusion events in degranulating mast cells.
    Nature. 1984 Nov 29-Dec 5;312(5993):453-5 PMID: 6504157
  15. Simultaneous electrical and optical measurements show that membrane fusion precedes secretory granule swelling during exocytosis of beige mouse mast cells.
    Proc Natl Acad Sci U S A. 1987 Mar;84(6):1585-9 PMID: 3470745
  16. Currents through the fusion pore that forms during exocytosis of a secretory vesicle.
    Nature. 1987 Aug 27-Sep 2;328(6133):814-7 PMID: 2442614
  17. Biomembrane fusion: a new concept derived from model studies using two interacting planar lipid bilayers.
    Biochim Biophys Acta. 1987 Oct 5;906(3):309-52 PMID: 3307918
  18. Cooperativity in viral fusion.
    Cell Biophys. 1988 Jan-Jun;12:1-12 PMID: 2453272
  19. Kinetics of pH-dependent fusion between 3T3 fibroblasts expressing influenza hemagglutinin and red blood cells. Measurement by dequenching of fluorescence.
    J Biol Chem. 1989 Mar 5;264(7):3972-8 PMID: 2917985
  20. Initial stages of influenza hemagglutinin-induced cell fusion monitored simultaneously by two fluorescent events: cytoplasmic continuity and lipid mixing.
    J Cell Biol. 1989 Jul;109(1):113-22 PMID: 2745545
  21. Patch clamp studies of single cell-fusion events mediated by a viral fusion protein.
    Nature. 1989 Nov 30;342(6249):555-8 PMID: 2586627
  22. On the use of self-quenching fluorophores in the study of membrane fusion kinetics. The effect of slow probe redistribution.
    Biophys Chem. 1989 Nov;34(3):283-92 PMID: 2611351
  23. Observation of single influenza virus-cell fusion and measurement by fluorescence video microscopy.
    Proc Natl Acad Sci U S A. 1990 Mar;87(5):1850-4 PMID: 2308945
  24. Real-time multi-wavelength fluorescence imaging of living cells.
    Biotechniques. 1990 Mar;8(3):296-308 PMID: 2331370
  25. Fusion of influenza hemagglutinin-expressing fibroblasts with glycophorin-bearing liposomes: role of hemagglutinin surface density.
    Biochemistry. 1990 Oct 16;29(41):9697-707 PMID: 2271610
  26. Delay time for influenza virus hemagglutinin-induced membrane fusion depends on hemagglutinin surface density.
    J Virol. 1991 May;65(5):2402-7 PMID: 1850019
  27. The first milliseconds of the pore formed by a fusogenic viral envelope protein during membrane fusion.
    Proc Natl Acad Sci U S A. 1991 May 1;88(9):3623-7 PMID: 2023911
  28. Single cell fusion events induced by influenza hemagglutinin: studies with rapid-flow, quantitative fluorescence microscopy.
    Exp Cell Res. 1991 Jul;195(1):137-44 PMID: 2055263
  29. A dissection of steps leading to viral envelope protein-mediated membrane fusion.
    Ann N Y Acad Sci. 1991;635:285-96 PMID: 1741588
  30. A long-lived state for influenza virus-erythrocyte complexes committed to fusion at neutral pH.
    FEBS Lett. 1992 Oct 26;311(3):221-5 PMID: 1397318
  31. The exocytotic fusion pore modeled as a lipidic pore.
    Biophys J. 1992 Oct;63(4):1118-32 PMID: 1420930
  32. The exocytotic fusion pore.
    J Cell Biol. 1992 Dec;119(6):1395-404 PMID: 1469040
  33. GPI-anchored influenza hemagglutinin induces hemifusion to both red blood cell and planar bilayer membranes.
    J Cell Biol. 1995 Nov;131(3):679-91 PMID: 7593189
  34. Membrane fusion mediated by the influenza virus hemagglutinin requires the concerted action of at least three hemagglutinin trimers.
    J Cell Biol. 1996 May;133(3):559-69 PMID: 8636231
  35. A quantitative description of membrane current and its application to conduction and excitation in nerve.
    J Physiol. 1952 Aug;117(4):500-44 PMID: 12991237
  36. Intermediates in influenza virus PR/8 haemagglutinin-induced membrane fusion.
    J Gen Virol. 1994 Feb;75 ( Pt 2):395-9 PMID: 8113761
  37. Influenza hemagglutinin-mediated fusion pores connecting cells to planar membranes: flickering to final expansion.
    J Gen Physiol. 1993 Dec;102(6):1131-49 PMID: 8133242
  38. Influenza virus hemagglutinin-induced cell-planar bilayer fusion: quantitative dissection of fusion pore kinetics into stages.
    J Gen Physiol. 1993 Dec;102(6):1151-70 PMID: 8133243
  39. The exocytotic fusion pore and neurotransmitter release.
    Neuron. 1994 Apr;12(4):707-16 PMID: 7909233
  40. Rapid simultaneous estimation of intracellular calcium and pH.
    Methods Cell Biol. 1994;40:183-220 PMID: 8201976
  41. Vesicle fusion from yeast to man.
    Nature. 1994 Jul 21;370(6486):191-3 PMID: 8028665
  42. Restricted movement of lipid and aqueous dyes through pores formed by influenza hemagglutinin during cell fusion.
    J Cell Biol. 1994 Dec;127(6 Pt 2):1885-94 PMID: 7806567
  43. Differences in dispersion of influenza virus lipids and proteins during fusion.
    Exp Cell Res. 1995 Feb;216(2):411-21 PMID: 7843286
  44. Transient domains induced by influenza haemagglutinin during membrane fusion.
    Mol Membr Biol. 1995 Jan-Mar;12(1):135-42 PMID: 7767373
  45. Membrane flux through the pore formed by a fusogenic viral envelope protein during cell fusion.
    J Cell Biol. 1993 May;121(3):543-52 PMID: 8486735
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.135.1.63
Email
[email protected]
Published
1996-10-00
Pages
63-71
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2121025
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.135.1.63 PMC Full Text View journal details
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