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PMID: 10637224 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Involvement of ezrin/moesin in de novo actin assembly on phagosomal membranes.

The EMBO journal ·Vol. 19 ·No. 2 ·2000-01-17 ·Pages 199-212

Defacque H, Egeberg M, Habermann A, Diakonova M, Roy C, Mangeat P, Voelter W, Marriott G, Pfannstiel J, Faulstich H, Griffiths G

Abstract

The current study focuses on the molecular mechanisms responsible for actin assembly on a defined membrane surface: the phagosome. Mature phagosomes were surrounded by filamentous actin in vivo in two different cell types. Fluorescence microscopy was used to study in vitro actin nucleation/polymerization (assembly) on the surface of phagosomes isolated from J774 mouse macrophages. In order to prevent non-specific actin polymerization during the assay, fluorescent G-actin was mixed with thymosin beta4. The cytoplasmic side of phagosomes induced de novo assembly and barbed end growth of actin filaments. This activity varied cyclically with the maturation state of phagosomes, both in vivo and in vitro. Peripheral membrane proteins are crucial components of this actin assembly machinery, and we demonstrate a role for ezrin and/or moesin in this process. We propose that this actin assembly process facilitates phagosome/endosome aggregation prior to membrane fusion.

MeSH Terms
Actins/biosynthesis,chemistry,metabolism Animals Cell Line Cytochalasin D/pharmacology Cytoskeletal Proteins Cytoskeleton/metabolism Intracellular Membranes/metabolism,ultrastructure Kinetics Macrophages Membrane Fusion Mice Microfilament Proteins/metabolism Microscopy, Immunoelectron Molecular Sequence Data Phagosomes/drug effects,metabolism,ultrastructure Phosphoproteins/metabolism Recombinant Proteins/metabolism Thymosin/metabolism Transfection
Chemicals
Actins Cytoskeletal Proteins Microfilament Proteins Phosphoproteins Recombinant Proteins ezrin moesin Cytochalasin D thymosin beta(4) Thymosin
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Defacque H
European Molecular Biology Laboratory, Meyerhofstrasse 1, Postfach 102209, 69012 Heidelberg, Germany.
Egeberg M
Habermann A
Diakonova M
Roy C
Mangeat P
Voelter W
Marriott G
Pfannstiel J
Faulstich H
Griffiths G
References (88)
88 references, click to expand
  1. How profilin promotes actin filament assembly in the presence of thymosin beta 4.
    Cell. 1993 Dec 3;75(5):1007-14 PMID: 8252614
  2. Actin filaments undergo limited subunit exchange in physiological salt conditions.
    J Cell Biol. 1982 Aug;94(2):316-24 PMID: 7202009
  3. The actin cytoskeleton is required for receptor-mediated endocytosis in mammalian cells.
    J Biol Chem. 1997 Aug 15;272(33):20332-5 PMID: 9252336
  4. Structure and function of the Arp2/3 complex.
    Curr Opin Struct Biol. 1999 Apr;9(2):244-9 PMID: 10322212
  5. Regulation of vinculin binding to talin and actin by phosphatidyl-inositol-4-5-bisphosphate.
    Nature. 1996 Jun 6;381(6582):531-5 PMID: 8632828
  6. Preparation of myosin and its subfragments from rabbit skeletal muscle.
    Methods Enzymol. 1982;85 Pt B:55-71 PMID: 6214692
  7. In vitro fusion of phagosomes with different endocytic organelles from J774 macrophages.
    J Biol Chem. 1998 Nov 13;273(46):30379-90 PMID: 9804802
  8. Ca(2+)-independent F-actin assembly and disassembly during Fc receptor-mediated phagocytosis in mouse macrophages.
    J Cell Biol. 1991 May;113(4):757-67 PMID: 2026648
  9. Radiolabeled latex particles ithe investigation of phagocytosis in vivo: clearance curves and histological observations.
    J Reticuloendothel Soc. 1969 Oct-Dec;6(5):561-89 PMID: 5349135
  10. Interaction of human Arp2/3 complex and the Listeria monocytogenes ActA protein in actin filament nucleation.
    Science. 1998 Jul 3;281(5373):105-8 PMID: 9651243
  11. Lysosomal enzyme trafficking between phagosomes, endosomes, and lysosomes in J774 macrophages. Enrichment of cathepsin H in early endosomes.
    J Biol Chem. 1998 Apr 17;273(16):9842-51 PMID: 9545324
  12. Capping protein terminates but does not initiate chemoattractant-induced actin assembly in Dictyostelium.
    J Cell Biol. 1997 Dec 1;139(5):1243-53 PMID: 9382870
  13. Microtubule-associated protein-dependent binding of phagosomes to microtubules.
    J Biol Chem. 1996 Feb 16;271(7):3803-11 PMID: 8631997
  14. Dynamics of the cytoskeleton in Amoeba proteus. I. Redistribution of microinjected fluorescein-labeled actin during locomotion, immobilization and phagocytosis.
    Cell Tissue Res. 1983;232(1):79-96 PMID: 6683994
  15. Mechanisms of actin rearrangements mediating platelet activation.
    J Cell Biol. 1992 Sep;118(6):1421-42 PMID: 1325975
  16. The biochemical basis of phagocytosis. I. Metabolic changes during the ingestion of particles by polymorphonuclear leukocytes.
    J Biol Chem. 1959 Jun;234(6):1355-62 PMID: 13654378
  17. Ezrin/radixin/moesin (ERM) proteins bind to a positively charged amino acid cluster in the juxta-membrane cytoplasmic domain of CD44, CD43, and ICAM-2.
    J Cell Biol. 1998 Feb 23;140(4):885-95 PMID: 9472040
  18. Subplasmalemmal microfilaments and microtubules in resting and phagocytizing cultivated macrophages.
    J Cell Biol. 1973 Oct;59(1):12-27 PMID: 4356569
  19. Acidic fibroblast growth factor is a modulator of epithelial plasticity in a rat bladder carcinoma cell line.
    Proc Natl Acad Sci U S A. 1990 Feb;87(3):1124-8 PMID: 2153969
  20. Control of actin dynamics.
    Curr Opin Cell Biol. 1998 Feb;10(1):45-51 PMID: 9484594
  21. Mechanism of action of cytochalasin: evidence that it binds to actin filament ends.
    J Cell Biol. 1981 Mar;88(3):487-91 PMID: 6894300
  22. ERM (ezrin/radixin/moesin) family: from cytoskeleton to signal transduction.
    Curr Opin Cell Biol. 1997 Feb;9(1):70-5 PMID: 9013673
  23. Actin dynamics in human neutrophils during adhesion and phagocytosis is controlled by changes in intracellular free calcium.
    Eur J Cell Biol. 1993 Oct;62(1):49-58 PMID: 8269978
  24. The carboxyl-terminal region of EBP50 binds to a site in the amino-terminal domain of ezrin that is masked in the dormant molecule.
    J Biol Chem. 1998 Jul 17;273(29):18452-8 PMID: 9660814
  25. Corequirement of specific phosphoinositides and small GTP-binding protein Cdc42 in inducing actin assembly in Xenopus egg extracts.
    J Cell Biol. 1998 Mar 9;140(5):1125-36 PMID: 9490725
  26. Thymosin beta 4 sequesters the majority of G-actin in resting human polymorphonuclear leukocytes.
    J Cell Biol. 1992 Dec;119(5):1261-70 PMID: 1447300
  27. Biogenesis of phagolysosomes proceeds through a sequential series of interactions with the endocytic apparatus.
    J Cell Biol. 1994 Mar;124(5):677-88 PMID: 8120091
  28. The ezrin protein family: membrane-cytoskeleton interactions and disease associations.
    Curr Opin Cell Biol. 1997 Oct;9(5):659-66 PMID: 9330869
  29. Differential expression of the human thymosin-beta 4 gene in lymphocytes, macrophages, and granulocytes.
    J Immunol. 1987 Dec 1;139(11):3840-8 PMID: 3500230
  30. Rapid phosphorylation and reorganization of ezrin and spectrin accompany morphological changes induced in A-431 cells by epidermal growth factor.
    J Cell Biol. 1989 Mar;108(3):921-30 PMID: 2646308
  31. Gaining insight into a complex organelle, the phagosome, using two-dimensional gel electrophoresis.
    Electrophoresis. 1995 Dec;16(12):2249-57 PMID: 8824784
  32. A contractile activity that closes phagosomes in macrophages.
    J Cell Sci. 1999 Feb;112 ( Pt 3):307-16 PMID: 9885284
  33. Regulation of cortical structure by the ezrin-radixin-moesin protein family.
    Curr Opin Cell Biol. 1999 Feb;11(1):109-16 PMID: 10047517
  34. Ezrin is an effector of hepatocyte growth factor-mediated migration and morphogenesis in epithelial cells.
    J Cell Biol. 1997 Jul 28;138(2):423-34 PMID: 9230083
  35. Association of ezrin with intercellular adhesion molecule-1 and -2 (ICAM-1 and ICAM-2). Regulation by phosphatidylinositol 4, 5-bisphosphate.
    J Biol Chem. 1998 Aug 21;273(34):21893-900 PMID: 9705328
  36. An actin-nucleating activity in polymorphonuclear leukocytes is modulated by chemotactic peptides.
    J Cell Biol. 1986 Dec;103(6 Pt 2):2707-14 PMID: 3793753
  37. Molecular characterization of phagosomes.
    J Biol Chem. 1994 Dec 23;269(51):32194-200 PMID: 7798218
  38. Biochemical characterization of ezrin-actin interaction.
    J Biol Chem. 1996 Mar 22;271(12):7224-9 PMID: 8636161
  39. Molecular requirements for bi-directional movement of phagosomes along microtubules.
    J Cell Biol. 1997 Apr 7;137(1):113-29 PMID: 9105041
  40. Peptide-specific antibodies localize the major lipid binding sites of talin dimers to oppositely arranged N-terminal 47 kDa subdomains.
    FEBS Lett. 1998 Apr 17;426(2):165-70 PMID: 9599000
  41. Phosphorylation of moesin by rho-associated kinase (Rho-kinase) plays a crucial role in the formation of microvilli-like structures.
    J Biol Chem. 1998 Dec 25;273(52):34663-6 PMID: 9856983
  42. Ezrin has properties to self-associate at the plasma membrane.
    J Cell Sci. 1994 Sep;107 ( Pt 9):2509-21 PMID: 7844168
  43. Rho- and rac-dependent assembly of focal adhesion complexes and actin filaments in permeabilized fibroblasts: an essential role for ezrin/radixin/moesin proteins.
    J Cell Biol. 1997 Aug 25;138(4):927-38 PMID: 9265657
  44. A role for MARCKS, the alpha isozyme of protein kinase C and myosin I in zymosan phagocytosis by macrophages.
    J Exp Med. 1995 Sep 1;182(3):829-40 PMID: 7650489
  45. Filamentous actin in Paramecium cells: mapping by phalloidin affinity labeling in vivo and in vitro.
    J Histochem Cytochem. 1986 Apr;34(4):443-54 PMID: 2419395
  46. Regulation of F-actin binding to platelet moesin in vitro by both phosphorylation of threonine 558 and polyphosphatidylinositides.
    Mol Biol Cell. 1999 Aug;10(8):2669-85 PMID: 10436021
  47. Interactions of Listeria monocytogenes with mammalian cells during entry and actin-based movement: bacterial factors, cellular ligands and signaling.
    EMBO J. 1998 Jul 15;17(14):3797-806 PMID: 9669997
  48. Actin-based cell motility and cell locomotion.
    Cell. 1996 Feb 9;84(3):371-9 PMID: 8608590
  49. Localization of five annexins in J774 macrophages and on isolated phagosomes.
    J Cell Sci. 1997 May;110 ( Pt 10):1199-213 PMID: 9191044
  50. Normal development of mice and unimpaired cell adhesion/cell motility/actin-based cytoskeleton without compensatory up-regulation of ezrin or radixin in moesin gene knockout.
    J Biol Chem. 1999 Jan 22;274(4):2315-21 PMID: 9890997
  51. Protein phosphorylation during phagosome maturation.
    FEBS Lett. 1996 Nov 25;398(1):37-42 PMID: 8946949
  52. Effects of cytochalasin and phalloidin on actin.
    J Cell Biol. 1987 Oct;105(4):1473-8 PMID: 3312229
  53. Ezrin contains cytoskeleton and membrane binding domains accounting for its proposed role as a membrane-cytoskeletal linker.
    J Cell Biol. 1993 Jan;120(1):129-39 PMID: 8416983
  54. The secretion-stimulated 80K phosphoprotein of parietal cells is ezrin, and has properties of a membrane cytoskeletal linker in the induced apical microvilli.
    EMBO J. 1991 Sep;10(9):2363-73 PMID: 1831124
  55. C-terminal threonine phosphorylation activates ERM proteins to link the cell's cortical lipid bilayer to the cytoskeleton.
    Biochem Biophys Res Commun. 1998 Dec 30;253(3):561-5 PMID: 9918767
  56. Identification of a phosphatidylinositol-4,5-bisphosphate-binding domain in the N-terminal region of ezrin.
    FEBS Lett. 1995 Dec 4;376(3):172-6 PMID: 7498535
  57. Structural relationships of actin-binding proteins.
    Curr Opin Cell Biol. 1992 Feb;4(1):36-42 PMID: 1558752
  58. Membrane-cytoskeleton interactions in animal cells.
    Biochim Biophys Acta. 1989 May 9;988(2):147-71 PMID: 2524216
  59. Professional and non-professional phagocytes: an introduction.
    Trends Cell Biol. 1995 Mar;5(3):85-7 PMID: 14732160
  60. Ezrin oligomers are major cytoskeletal components of placental microvilli: a proposal for their involvement in cortical morphogenesis.
    J Cell Biol. 1995 Dec;131(5):1231-42 PMID: 8522586
  61. Moesin, the major ERM protein of lymphocytes and platelets, differs from ezrin in its insensitivity to calpain.
    FEBS Lett. 1999 Jan 22;443(1):31-6 PMID: 9928947
  62. RhoA-dependent phosphorylation and relocalization of ERM proteins into apical membrane/actin protrusions in fibroblasts.
    Mol Biol Cell. 1998 Feb;9(2):403-19 PMID: 9450964
  63. Actin polymerization and pseudopod extension during amoeboid chemotaxis.
    Cell Motil Cytoskeleton. 1988;10(1-2):77-90 PMID: 3052871
  64. Direct interaction of the Rho GDP dissociation inhibitor with ezrin/radixin/moesin initiates the activation of the Rho small G protein.
    J Biol Chem. 1997 Sep 12;272(37):23371-5 PMID: 9287351
  65. Identification of two distinct mechanisms of phagocytosis controlled by different Rho GTPases.
    Science. 1998 Nov 27;282(5394):1717-21 PMID: 9831565
  66. Actin and actin-binding proteins. A critical evaluation of mechanisms and functions.
    Annu Rev Biochem. 1986;55:987-1035 PMID: 3527055
  67. Actin filaments and the growth, movement, and spread of the intracellular bacterial parasite, Listeria monocytogenes.
    J Cell Biol. 1989 Oct;109(4 Pt 1):1597-608 PMID: 2507553
  68. Arp2/3 complex and actin depolymerizing factor/cofilin in dendritic organization and treadmilling of actin filament array in lamellipodia.
    J Cell Biol. 1999 May 31;145(5):1009-26 PMID: 10352018
  69. Fluorimetry study of N-(1-pyrenyl)iodoacetamide-labelled F-actin. Local structural change of actin protomer both on polymerization and on binding of heavy meromyosin.
    Eur J Biochem. 1981;114(1):33-8 PMID: 7011802
  70. Actin filament disassembly is a sufficient final trigger for exocytosis in nonexcitable cells.
    J Cell Biol. 1995 Feb;128(4):589-98 PMID: 7860632
  71. Regulation of actin polymerization in cell-free systems by GTPgammaS and Cdc42.
    J Cell Biol. 1997 Jul 28;138(2):363-74 PMID: 9230078
  72. Novel forms of epithelial cell motility on collagen and on glass surfaces.
    Cell Motil. 1982;2(4):333-41 PMID: 7151140
  73. Regulation mechanism of ERM (ezrin/radixin/moesin) protein/plasma membrane association: possible involvement of phosphatidylinositol turnover and Rho-dependent signaling pathway.
    J Cell Biol. 1996 Oct;135(1):37-51 PMID: 8858161
  74. Dephosphorylation of ezrin as an early event in renal microvillar breakdown and anoxic injury.
    Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7495-9 PMID: 7638219
  75. A dual involvement of the amino-terminal domain of ezrin in F- and G-actin binding.
    J Biol Chem. 1997 Aug 8;272(32):20088-95 PMID: 9242682
  76. Microfilament dynamics during cell movement and chemotaxis monitored using a GFP-actin fusion protein.
    Curr Biol. 1997 Mar 1;7(3):176-83 PMID: 9276758
  77. The suitability and application of a GFP-actin fusion protein for long-term imaging of the organization and dynamics of the cytoskeleton in mammalian cells.
    Eur J Cell Biol. 1998 Oct;77(2):81-90 PMID: 9840457
  78. Dictyostelium discoideum plasma membranes contain an actin-nucleating activity that requires ponticulin, an integral membrane glycoprotein.
    J Cell Biol. 1990 Mar;110(3):681-92 PMID: 2307703
  79. Actin filament organization in the fish keratocyte lamellipodium.
    J Cell Biol. 1995 Jun;129(5):1275-86 PMID: 7775574
  80. Interaction of thymosin beta 4 with muscle and platelet actin: implications for actin sequestration in resting platelets.
    Biochemistry. 1992 Jul 14;31(27):6179-85 PMID: 1627561
  81. Interaction of cytoskeletal proteins with membrane lipids.
    Int Rev Cytol. 1998;178:73-125 PMID: 9348669
  82. The kinetics of chemotactic peptide-induced change in F-actin content, F-actin distribution, and the shape of neutrophils.
    J Cell Biol. 1985 Sep;101(3):1078-85 PMID: 4040915
  83. Transport of phagosomes in mouse peritoneal macrophages.
    J Cell Sci. 1989 Sep;94 ( Pt 1):143-53 PMID: 2693471
  84. Rho-kinase phosphorylates COOH-terminal threonines of ezrin/radixin/moesin (ERM) proteins and regulates their head-to-tail association.
    J Cell Biol. 1998 Feb 9;140(3):647-57 PMID: 9456324
  85. Measurement of actin polymerization and cross-linking in agonist-stimulated cells.
    Methods Enzymol. 1991;196:486-96 PMID: 1851942
  86. Phagosomal proteins of Dictyostelium discoideum.
    J Eukaryot Microbiol. 1997 Jul-Aug;44(4):284-92 PMID: 9225442
  87. ERM proteins in cell adhesion and membrane dynamics.
    Trends Cell Biol. 1999 May;9(5):187-92 PMID: 10322453
  88. Separation of subfragment-1 isoenzymes from rabbit skeletal muscle myosin.
    Nature. 1975 Sep 4;257(5521):54-6 PMID: 125854
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/19.2.199
Published
2000-01-17
Pages
199-212
Language
English
Region
England
NLM ID
8208664
PMCID
PMC305554
Subset
IM
Databases
PDB
1D4B
External Links
PubMed source DOI: 10.1093/emboj/19.2.199 PMC Full Text View journal details
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