Home LiteratureArticle Details
PMID: 11018051 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cortactin localization to sites of actin assembly in lamellipodia requires interactions with F-actin and the Arp2/3 complex.

The Journal of cell biology ·Vol. 151 ·No. 1 ·2000-10-02 ·Pages 29-40

Weed SA, Karginov AV, Schafer DA, Weaver AM, Kinley AW, Cooper JA, Parsons JT

Abstract

Cortactin is an actin-binding protein that is enriched within the lamellipodia of motile cells and in neuronal growth cones. Here, we report that cortactin is localized with the actin-related protein (Arp) 2/3 complex at sites of actin polymerization within the lamellipodia. Two distinct sequence motifs of cortactin contribute to its interaction with the cortical actin network: the fourth of six tandem repeats and the amino-terminal acidic region (NTA). Cortactin variants lacking either the fourth tandem repeat or the NTA failed to localize at the cell periphery. Tandem repeat four was necessary for cortactin to stably bind F-actin in vitro. The NTA region interacts directly with the Arp2/3 complex based on affinity chromatography, immunoprecipitation assays, and binding assays using purified components. Cortactin variants containing the NTA region were inefficient at promoting Arp2/3 actin nucleation activity. These data provide strong evidence that cortactin is specifically localized to sites of dynamic cortical actin assembly via simultaneous interaction with F-actin and the Arp2/3 complex. Cortactin interacts via its Src homology 3 (SH3) domain with ZO-1 and the SHANK family of postsynaptic density 95/dlg/ZO-1 homology (PDZ) domain-containing proteins, suggesting that cortactin contributes to the spatial organization of sites of actin polymerization coupled to selected cell surface transmembrane receptor complexes.

MeSH Terms
Actin-Related Protein 2 Actin-Related Protein 3 Actins/metabolism Amino Acid Sequence Animals Binding Sites Biological Transport Cells, Cultured Cortactin Cytoskeletal Proteins Mice Microfilament Proteins/metabolism Molecular Sequence Data Protein Binding Protein Structure, Tertiary Pseudopodia Repetitive Sequences, Amino Acid Sequence Homology, Amino Acid rac GTP-Binding Proteins/metabolism
Chemicals
Actin-Related Protein 2 Actin-Related Protein 3 Actins Actr2 protein, mouse Actr3 protein, mouse Cortactin Cttn protein, mouse Cytoskeletal Proteins Microfilament Proteins rac GTP-Binding Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Weed S A
Department of Microbiology and Cancer Center, University of Virginia Health Sciences Center, Charlottesville, Virginia 22908, USA.
Karginov A V
Schafer D A
Weaver A M
Kinley A W
Cooper J A
Parsons J T
References (96)
96 references, click to expand
  1. Murine cortactin is phosphorylated in response to fibroblast growth factor-1 on tyrosine residues late in the G1 phase of the BALB/c 3T3 cell cycle.
    J Biol Chem. 1993 Nov 15;268(32):24427-31 PMID: 7693700
  2. The cytoskeleton in development of epithelial cell polarity.
    J Cell Sci Suppl. 1993;17:65-73 PMID: 7511618
  3. Complete DNA sequence of yeast chromosome XI.
    Nature. 1994 Jun 2;369(6479):371-8 PMID: 8196765
  4. The protein tyrosine kinase substrate cortactin is differentially expressed in murine B lymphoid tumors.
    Oncogene. 1994 Jul;9(7):1989-97 PMID: 7516062
  5. Direct association of occludin with ZO-1 and its possible involvement in the localization of occludin at tight junctions.
    J Cell Biol. 1994 Dec;127(6 Pt 1):1617-26 PMID: 7798316
  6. The Ras-related protein Cdc42Hs and bradykinin promote formation of peripheral actin microspikes and filopodia in Swiss 3T3 fibroblasts.
    Mol Cell Biol. 1995 Apr;15(4):1942-52 PMID: 7891688
  7. Rho, rac, and cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia.
    Cell. 1995 Apr 7;81(1):53-62 PMID: 7536630
  8. Invasion of epithelial cells by Shigella flexneri induces tyrosine phosphorylation of cortactin by a pp60c-src-mediated signalling pathway.
    EMBO J. 1995 Jun 1;14(11):2471-82 PMID: 7540134
  9. Tyrosine phosphorylation of p130Cas and cortactin accompanies integrin-mediated cell adhesion to extracellular matrix.
    J Biol Chem. 1995 Sep 22;270(38):22259-62 PMID: 7545676
  10. The mouse homolog of the Wiskott-Aldrich syndrome protein (WASP) gene is highly conserved and maps near the scurfy (sf) mutation on the X chromosome.
    Genomics. 1995 Sep 20;29(2):471-7 PMID: 8666397
  11. Amplification and expression of EMS-1 (cortactin) in head and neck squamous cell carcinoma cell lines.
    Oncogene. 1996 Jan 4;12(1):31-5 PMID: 8552396
  12. Actin-based cell motility and cell locomotion.
    Cell. 1996 Feb 9;84(3):371-9 PMID: 8608590
  13. Signal transduction and actin filament organization.
    Curr Opin Cell Biol. 1996 Feb;8(1):66-73 PMID: 8791404
  14. Rac and Cdc42 induce actin polymerization and G1 cell cycle progression independently of p65PAK and the JNK/SAPK MAP kinase cascade.
    Cell. 1996 Nov 1;87(3):519-29 PMID: 8898204
  15. Faciogenital dysplasia protein (FGD1) and Vav, two related proteins required for normal embryonic development, are upstream regulators of Rho GTPases.
    Curr Biol. 1996 Dec 1;6(12):1628-33 PMID: 8994827
  16. Actin polymerization is induced by Arp2/3 protein complex at the surface of Listeria monocytogenes.
    Nature. 1997 Jan 16;385(6613):265-9 PMID: 9000076
  17. Role of actin in the organisation of brain postsynaptic densities.
    Brain Res Mol Brain Res. 1996 Dec 31;43(1-2):246-50 PMID: 9037539
  18. Regulation of cell motility by mitogen-activated protein kinase.
    J Cell Biol. 1997 Apr 21;137(2):481-92 PMID: 9128257
  19. Identification of two regions in the N-terminal domain of ActA involved in the actin comet tail formation by Listeria monocytogenes.
    EMBO J. 1997 Apr 1;16(7):1531-40 PMID: 9130698
  20. Down-regulation of the filamentous actin cross-linking activity of cortactin by Src-mediated tyrosine phosphorylation.
    J Biol Chem. 1997 May 23;272(21):13911-5 PMID: 9153252
  21. Tight junction assembly during mouse blastocyst formation is regulated by late expression of ZO-1 alpha+ isoform.
    Development. 1997 May;124(10):2027-37 PMID: 9169849
  22. Translocation of cortactin to the cell periphery is mediated by the small GTPase Rac1.
    J Cell Sci. 1998 Aug;111 ( Pt 16):2433-43 PMID: 9683637
  23. SCAR, a WASP-related protein, isolated as a suppressor of receptor defects in late Dictyostelium development.
    J Cell Biol. 1998 Sep 7;142(5):1325-35 PMID: 9732292
  24. Cortactin localization in actin-containing adult and fetal tissues.
    J Histochem Cytochem. 1998 Oct;46(10):1189-91 PMID: 9742075
  25. Identification of a novel cortactin SH3 domain-binding protein and its localization to growth cones of cultured neurons.
    Mol Cell Biol. 1998 Oct;18(10):5838-51 PMID: 9742101
  26. The role of tyrosine phosphorylation of cortactin in the locomotion of endothelial cells.
    J Biol Chem. 1998 Oct 2;273(40):25770-6 PMID: 9748248
  27. A single gene product, claudin-1 or -2, reconstitutes tight junction strands and recruits occludin in fibroblasts.
    J Cell Biol. 1998 Oct 19;143(2):391-401 PMID: 9786950
  28. Cortactin associates with the cell-cell junction protein ZO-1 in both Drosophila and mouse.
    J Biol Chem. 1998 Nov 6;273(45):29672-7 PMID: 9792678
  29. The tight junction protein ZO-1 establishes a link between the transmembrane protein occludin and the actin cytoskeleton.
    J Biol Chem. 1998 Nov 6;273(45):29745-53 PMID: 9792688
  30. Isolation and characterization of cortactin isoforms and a novel cortactin-binding protein, CBP90.
    Genes Cells. 1998 Sep;3(9):603-12 PMID: 9813110
  31. WAVE, a novel WASP-family protein involved in actin reorganization induced by Rac.
    EMBO J. 1998 Dec 1;17(23):6932-41 PMID: 9843499
  32. Epidermal growth factor receptor-mediated motility in fibroblasts.
    Microsc Res Tech. 1998 Dec 1;43(5):395-411 PMID: 9858337
  33. Ribotypes and virulence gene polymorphisms suggest three distinct Listeria monocytogenes lineages with differences in pathogenic potential.
    Infect Immun. 1997 Jul;65(7):2707-16 PMID: 9199440
  34. The human Arp2/3 complex is composed of evolutionarily conserved subunits and is localized to cellular regions of dynamic actin filament assembly.
    J Cell Biol. 1997 Jul 28;138(2):375-84 PMID: 9230079
  35. Role of actin polymerization and adhesion to extracellular matrix in Rac- and Rho-induced cytoskeletal reorganization.
    J Cell Biol. 1997 Aug 25;138(4):913-26 PMID: 9265656
  36. Identification of N-WASP homologs in human and rat brain.
    Gene. 1997 Sep 1;196(1-2):43-8 PMID: 9322739
  37. Na,K-ATPase transport from endoplasmic reticulum to Golgi requires the Golgi spectrin-ankyrin G119 skeleton in Madin Darby canine kidney cells.
    Proc Natl Acad Sci U S A. 1997 Sep 30;94(20):10711-6 PMID: 9380700
  38. Mammalian actin-related protein 2/3 complex localizes to regions of lamellipodial protrusion and is composed of evolutionarily conserved proteins.
    Biochem J. 1997 Nov 15;328 ( Pt 1):105-12 PMID: 9359840
  39. Association of cortactin with developing neuromuscular specializations.
    J Neurocytol. 1997 Oct;26(10):637-50 PMID: 9368878
  40. Infection with Chlamydia trachomatis alters the tyrosine phosphorylation and/or localization of several host cell proteins including cortactin.
    Infect Immun. 1997 Dec;65(12):5301-8 PMID: 9393830
  41. EGF stimulates an increase in actin nucleation and filament number at the leading edge of the lamellipod in mammary adenocarcinoma cells.
    J Cell Sci. 1998 Jan;111 ( Pt 2):199-211 PMID: 9405304
  42. p140Sra-1 (specifically Rac1-associated protein) is a novel specific target for Rac1 small GTPase.
    J Biol Chem. 1998 Jan 2;273(1):291-5 PMID: 9417078
  43. Induction of filopodium formation by a WASP-related actin-depolymerizing protein N-WASP.
    Nature. 1998 Jan 1;391(6662):93-6 PMID: 9422512
  44. Rho GTPases and the actin cytoskeleton.
    Science. 1998 Jan 23;279(5350):509-14 PMID: 9438836
  45. In vivo functions of actin-binding proteins.
    Curr Opin Cell Biol. 1998 Feb;10(1):102-11 PMID: 9484601
  46. Cortactin-Src kinase signaling pathway is involved in N-syndecan-dependent neurite outgrowth.
    J Biol Chem. 1998 Apr 24;273(17):10702-8 PMID: 9553134
  47. The interaction of Arp2/3 complex with actin: nucleation, high affinity pointed end capping, and formation of branching networks of filaments.
    Proc Natl Acad Sci U S A. 1998 May 26;95(11):6181-6 PMID: 9600938
  48. Role of phosphatidylinositol 4,5-bisphosphate in Ras/Rac-induced disruption of the cortactin-actomyosin II complex and malignant transformation.
    Mol Cell Biol. 1998 Jul;18(7):3829-37 PMID: 9632767
  49. Database searching using mass spectrometry data.
    Electrophoresis. 1998 May;19(6):893-900 PMID: 9638935
  50. Scar, a WASp-related protein, activates nucleation of actin filaments by the Arp2/3 complex.
    Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):3739-44 PMID: 10097107
  51. Bidirectional signaling between the cytoskeleton and integrins.
    Curr Opin Cell Biol. 1999 Apr;11(2):274-86 PMID: 10209151
  52. The interaction between N-WASP and the Arp2/3 complex links Cdc42-dependent signals to actin assembly.
    Cell. 1999 Apr 16;97(2):221-31 PMID: 10219243
  53. 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
  54. Agonist-dependent interaction of the rat somatostatin receptor subtype 2 with cortactin-binding protein 1.
    J Biol Chem. 1999 Jun 25;274(26):18153-6 PMID: 10373412
  55. Specialized surface protrusions of invasive cells, invadopodia and lamellipodia, have differential MT1-MMP, MMP-2, and TIMP-2 localization.
    Ann N Y Acad Sci. 1999 Jun 30;878:361-71 PMID: 10415741
  56. Shank, a novel family of postsynaptic density proteins that binds to the NMDA receptor/PSD-95/GKAP complex and cortactin.
    Neuron. 1999 Jul;23(3):569-82 PMID: 10433268
  57. Coupling of mGluR/Homer and PSD-95 complexes by the Shank family of postsynaptic density proteins.
    Neuron. 1999 Jul;23(3):583-92 PMID: 10433269
  58. An invasion-related complex of cortactin, paxillin and PKCmu associates with invadopodia at sites of extracellular matrix degradation.
    Oncogene. 1999 Aug 5;18(31):4440-9 PMID: 10442635
  59. Coronin localizes to leading edges and is involved in cell spreading and lamellipodium extension in vertebrate cells.
    J Cell Sci. 1999 Sep;112 (Pt 17):2833-42 PMID: 10444378
  60. Activation of the CDC42 effector N-WASP by the Shigella flexneri IcsA protein promotes actin nucleation by Arp2/3 complex and bacterial actin-based motility.
    J Cell Biol. 1999 Sep 20;146(6):1319-32 PMID: 10491394
  61. Proline-rich synapse-associated proteins ProSAP1 and ProSAP2 interact with synaptic proteins of the SAPAP/GKAP family.
    Biochem Biophys Res Commun. 1999 Oct 14;264(1):247-52 PMID: 10527873
  62. FSP95, a testis-specific 95-kilodalton fibrous sheath antigen that undergoes tyrosine phosphorylation in capacitated human spermatozoa.
    Biol Reprod. 1999 Nov;61(5):1184-97 PMID: 10529264
  63. Regulation of actin polymerization by Arp2/3 complex and WASp/Scar proteins.
    J Biol Chem. 1999 Nov 12;274(46):32531-4 PMID: 10551802
  64. Influence of the C terminus of Wiskott-Aldrich syndrome protein (WASp) and the Arp2/3 complex on actin polymerization.
    Biochemistry. 1999 Nov 16;38(46):15212-22 PMID: 10563804
  65. Spatial control of actin polymerization during neutrophil chemotaxis.
    Nat Cell Biol. 1999 Jun;1(2):75-81 PMID: 10559877
  66. Src is required for cell migration and shape changes induced by fibroblast growth factor 1.
    Oncogene. 1999 Nov 18;18(48):6700-6 PMID: 10597276
  67. Direct binding of three tight junction-associated MAGUKs, ZO-1, ZO-2, and ZO-3, with the COOH termini of claudins.
    J Cell Biol. 1999 Dec 13;147(6):1351-63 PMID: 10601346
  68. Potentiation of cell migration by adhesion-dependent cooperative signals from the GTPase Rac and Raf kinase.
    J Biol Chem. 1999 Dec 31;274(53):37855-61 PMID: 10608850
  69. Association of mouse actin-binding protein 1 (mAbp1/SH3P7), an Src kinase target, with dynamic regions of the cortical actin cytoskeleton in response to Rac1 activation.
    Mol Biol Cell. 2000 Jan;11(1):393-412 PMID: 10637315
  70. A role for myosin-I in actin assembly through interactions with Vrp1p, Bee1p, and the Arp2/3 complex.
    J Cell Biol. 2000 Jan 24;148(2):353-62 PMID: 10648568
  71. Direct involvement of yeast type I myosins in Cdc42-dependent actin polymerization.
    J Cell Biol. 2000 Jan 24;148(2):363-73 PMID: 10648569
  72. Polarization of chemoattractant receptor signaling during neutrophil chemotaxis.
    Science. 2000 Feb 11;287(5455):1037-40 PMID: 10669415
  73. Actin machinery: pushing the envelope.
    Curr Opin Cell Biol. 2000 Feb;12(1):104-12 PMID: 10679366
  74. Pyrene actin: documentation of the validity of a sensitive assay for actin polymerization.
    J Muscle Res Cell Motil. 1983 Apr;4(2):253-62 PMID: 6863518
  75. Identification of ZO-1: a high molecular weight polypeptide associated with the tight junction (zonula occludens) in a variety of epithelia.
    J Cell Biol. 1986 Sep;103(3):755-66 PMID: 3528172
  76. Contact formation during fibroblast locomotion: involvement of membrane ruffles and microtubules.
    J Cell Biol. 1988 Mar;106(3):747-60 PMID: 3126193
  77. An unusual beta-spectrin associated with clustered acetylcholine receptors.
    J Cell Biol. 1989 Feb;108(2):481-93 PMID: 2645300
  78. Isolation and characterization of a novel human gene expressed specifically in the cells of hematopoietic lineage.
    Nucleic Acids Res. 1989 Nov 25;17(22):9367-79 PMID: 2587259
  79. Monoclonal antibodies to individual tyrosine-phosphorylated protein substrates of oncogene-encoded tyrosine kinases.
    Proc Natl Acad Sci U S A. 1990 May;87(9):3328-32 PMID: 2110361
  80. Identification and characterization of a novel cytoskeleton-associated pp60src substrate.
    Mol Cell Biol. 1991 Oct;11(10):5113-24 PMID: 1922035
  81. The small GTP-binding protein rac regulates growth factor-induced membrane ruffling.
    Cell. 1992 Aug 7;70(3):401-10 PMID: 1643658
  82. Activation of the MAP kinase pathway by FGF-1 correlates with cell proliferation induction while activation of the Src pathway correlates with migration.
    J Cell Biol. 1998 Jun 29;141(7):1647-58 PMID: 9647656
  83. 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
  84. Structural and functional regulation of tight junctions by RhoA and Rac1 small GTPases.
    J Cell Biol. 1998 Jul 13;142(1):101-15 PMID: 9660866
  85. Overexpression of EMS1/cortactin in NIH3T3 fibroblasts causes increased cell motility and invasion in vitro.
    Oncogene. 1998 Jun 25;16(25):3227-32 PMID: 9681820
  86. Visualization and molecular analysis of actin assembly in living cells.
    J Cell Biol. 1998 Dec 28;143(7):1919-30 PMID: 9864364
  87. Scar1 and the related Wiskott-Aldrich syndrome protein, WASP, regulate the actin cytoskeleton through the Arp2/3 complex.
    Curr Biol. 1998 Dec 17-31;8(25):1347-56 PMID: 9889097
  88. Overcoming expression and purification problems of RhoGDI using a family of "parallel" expression vectors.
    Protein Expr Purif. 1999 Feb;15(1):34-9 PMID: 10024467
  89. Functional design in the actin cytoskeleton.
    Curr Opin Cell Biol. 1999 Feb;11(1):54-60 PMID: 10047522
  90. The Arp2/3 complex: a multifunctional actin organizer.
    Curr Opin Cell Biol. 1999 Feb;11(1):117-21 PMID: 10047519
  91. Cell shrinkage regulates Src kinases and induces tyrosine phosphorylation of cortactin, independent of the osmotic regulation of Na+/H+ exchangers.
    J Biol Chem. 1999 Mar 19;274(12):8093-102 PMID: 10075710
  92. Actin polymerization: Where the WASP stings.
    Curr Biol. 1999 Mar 11;9(5):R160-3 PMID: 10074445
  93. Amplification of genes within the chromosome 11q13 region is indicative of poor prognosis in patients with operable breast cancer.
    Cancer Res. 1992 Oct 1;52(19):5229-34 PMID: 1394126
  94. Identification and characterization of a cytoskeleton-associated, epidermal growth factor sensitive pp60c-src substrate.
    Oncogene. 1992 Dec;7(12):2429-38 PMID: 1281304
  95. Cortactin, an 80/85-kilodalton pp60src substrate, is a filamentous actin-binding protein enriched in the cell cortex.
    J Cell Biol. 1993 Mar;120(6):1417-26 PMID: 7680654
  96. The 220-kD protein colocalizing with cadherins in non-epithelial cells is identical to ZO-1, a tight junction-associated protein in epithelial cells: cDNA cloning and immunoelectron microscopy.
    J Cell Biol. 1993 May;121(3):491-502 PMID: 8486731
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.151.1.29
Published
2000-10-02
Pages
29-40
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2189811
Subset
IM
Grants
NCI NIH HHS · CA29243 · United States
NIGMS NIH HHS · R01 GM038542 · United States
NIGMS NIH HHS · R01 GM038542-11S1 · United States
NIGMS NIH HHS · GM38542 · United States
NCI NIH HHS · CA40042 · United States
External Links
PubMed source DOI: 10.1083/jcb.151.1.29 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2