Home LiteratureArticle Details
PMID: 10970850 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Ezrin function is required for ROCK-mediated fibroblast transformation by the Net and Dbl oncogenes.

The EMBO journal ·Vol. 19 ·No. 17 ·2000-09-01 ·Pages 4565-76

Tran Quang C, Gautreau A, Arpin M, Treisman R

Abstract

The small G protein RhoA and its GDP/GTP exchange factors (GEFs) Net and Dbl can transform NIH 3T3 fibroblasts, dependent on the activity of the RhoA effector kinase ROCK. We investigated the role of the cytoskeletal linker protein ezrin in this process. RhoA effector loop mutants which can bind ROCK induce relocalization of ezrin to dorsal actin-containing cell surface protrusions, as do Net and Dbl. Both processes are inhibited by the ROCK inhibitor Y27632, which also inhibits association of ezrin with the cytoskeleton, and phosphorylation of T567, conserved between ezrin and its relatives radixin and moesin. ROCK can phosphorylate the ezrin C-terminus in vitro. The ezrin mutant T567A cannot be relocalized by activated RhoA, Net or Dbl or by ROCK itself, and also inhibits RhoA-mediated contractility and focal adhesion formation. Moreover, ezrin T567A, but not wild-type ezrin, restores contact inhibition to Net- and Dbl-transformed cells, and inhibits the activity of Net and Ras in focus formation assays. These results implicate ROCK-mediated ezrin C-terminal phosphorylation in transformation by RhoGEFs.

MeSH Terms
3T3 Cells Animals Cell Transformation, Neoplastic/genetics Cytoskeletal Proteins Cytoskeleton/enzymology Guanine Nucleotide Exchange Factors Intracellular Signaling Peptides and Proteins Mice Oncogene Proteins Oncogenes Phosphoproteins/chemistry,metabolism,physiology Phosphorylation Protein Serine-Threonine Kinases/physiology Proto-Oncogene Proteins c-ets Retroviridae Proteins, Oncogenic/genetics Signal Transduction Transcription Factors/genetics rho-Associated Kinases
Chemicals
Cytoskeletal Proteins Elk3 protein, mouse Guanine Nucleotide Exchange Factors Intracellular Signaling Peptides and Proteins Mcf2 protein, mouse Oncogene Proteins Phosphoproteins Proto-Oncogene Proteins c-ets Retroviridae Proteins, Oncogenic Transcription Factors ezrin Protein Serine-Threonine Kinases rho-Associated Kinases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Tran Quang C
Transcription Laboratory, Room 528, Imperial Cancer Research Fund Laboratories, 44 Lincoln's Inn Fields, London WC2A 3PX, UK.
Gautreau A
Arpin M
Treisman R
References (53)
53 references, click to expand
  1. An essential role for Rho, Rac, and Cdc42 GTPases in cell cycle progression through G1.
    Science. 1995 Sep 1;269(5228):1270-2 PMID: 7652575
  2. Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension.
    Nature. 1997 Oct 30;389(6654):990-4 PMID: 9353125
  3. Activation of Rac1, RhoA, and mitogen-activated protein kinases is required for Ras transformation.
    Mol Cell Biol. 1995 Nov;15(11):6443-53 PMID: 7565796
  4. 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
  5. A role for Rho in Ras transformation.
    Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11781-5 PMID: 8524848
  6. Activation of ERM proteins in vivo by Rho involves phosphatidyl-inositol 4-phosphate 5-kinase and not ROCK kinases.
    Curr Biol. 1999 Nov 4;9(21):1259-62 PMID: 10556088
  7. Cooperation between mDia1 and ROCK in Rho-induced actin reorganization.
    Nat Cell Biol. 1999 Jul;1(3):136-43 PMID: 10559899
  8. Ezrin, a membrane-cytoskeletal linking protein, is involved in the process of invasion of endometrial cancer cells.
    Cancer Lett. 1999 Dec 1;147(1-2):31-8 PMID: 10660086
  9. Stimulation of phosphatidylinositol-4-phosphate 5-kinase by Rho-kinase.
    J Biol Chem. 2000 Apr 7;275(14):10168-74 PMID: 10744700
  10. The TSC1 tumour suppressor hamartin regulates cell adhesion through ERM proteins and the GTPase Rho.
    Nat Cell Biol. 2000 May;2(5):281-7 PMID: 10806479
  11. Structure of the ERM protein moesin reveals the FERM domain fold masked by an extended actin binding tail domain.
    Cell. 2000 Apr 28;101(3):259-70 PMID: 10847681
  12. Morphogenic effects of ezrin require a phosphorylation-induced transition from oligomers to monomers at the plasma membrane.
    J Cell Biol. 2000 Jul 10;150(1):193-203 PMID: 10893267
  13. 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
  14. ERM family members as molecular linkers between the cell surface glycoprotein CD44 and actin-based cytoskeletons.
    J Cell Biol. 1994 Jul;126(2):391-401 PMID: 7518464
  15. An anti-Ras function of neurofibromatosis type 2 gene product (NF2/Merlin).
    J Biol Chem. 1994 Sep 23;269(38):23387-90 PMID: 8089100
  16. Dbl and Vav mediate transformation via mitogen-activated protein kinase pathways that are distinct from those activated by oncogenic Ras.
    Mol Cell Biol. 1994 Oct;14(10):6848-57 PMID: 7935402
  17. Deregulation of genes encoding microfilament-associated proteins during Fos-induced morphological transformation.
    Oncogene. 1995 Feb 2;10(3):603-8 PMID: 7845686
  18. Soluble ezrin purified from placenta exists as stable monomers and elongated dimers with masked C-terminal ezrin-radixin-moesin association domains.
    Biochemistry. 1995 Dec 26;34(51):16830-7 PMID: 8527459
  19. Phosphorylation of threonine 558 in the carboxyl-terminal actin-binding domain of moesin by thrombin activation of human platelets.
    J Biol Chem. 1995 Dec 29;270(52):31377-85 PMID: 8537411
  20. Regulation of myosin phosphatase by Rho and Rho-associated kinase (Rho-kinase)
    Science. 1996 Jul 12;273(5272):245-8 PMID: 8662509
  21. A highly expressed 81 kDa protein in immortalized mouse fibroblast: its proliferative function and identity with ezrin.
    Oncogene. 1996 Sep 19;13(6):1231-7 PMID: 8808697
  22. The p160 RhoA-binding kinase ROK alpha is a member of a kinase family and is involved in the reorganization of the cytoskeleton.
    Mol Cell Biol. 1996 Oct;16(10):5313-27 PMID: 8816443
  23. Transformation by Rho exchange factor oncogenes is mediated by activation of an integrin-dependent pathway.
    EMBO J. 1996 Dec 2;15(23):6525-30 PMID: 8978679
  24. Formation of actin stress fibers and focal adhesions enhanced by Rho-kinase.
    Science. 1997 Feb 28;275(5304):1308-11 PMID: 9036856
  25. Dbl family proteins.
    Biochim Biophys Acta. 1997 Feb 22;1332(1):F1-23 PMID: 9061011
  26. Massive actin polymerization induced by phosphatidylinositol-4-phosphate 5-kinase in vivo.
    J Biol Chem. 1997 Mar 21;272(12):7578-81 PMID: 9065410
  27. p160ROCK, a Rho-associated coiled-coil forming protein kinase, works downstream of Rho and induces focal adhesions.
    FEBS Lett. 1997 Mar 10;404(2-3):118-24 PMID: 9119047
  28. Ezrin: a protein requiring conformational activation to link microfilaments to the plasma membrane in the assembly of cell surface structures.
    J Cell Sci. 1997 Dec;110 ( Pt 24):3011-8 PMID: 9365271
  29. Ras-stimulated extracellular signal-related kinase 1 and RhoA activities coordinate platelet-derived growth factor-induced G1 progression through the independent regulation of cyclin D1 and p27.
    J Biol Chem. 1997 Dec 26;272(52):32966-71 PMID: 9407076
  30. 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
  31. 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
  32. RhoA effector mutants reveal distinct effector pathways for cytoskeletal reorganization, SRF activation and transformation.
    EMBO J. 1998 Mar 2;17(5):1350-61 PMID: 9482732
  33. Protein kinase C-theta phosphorylation of moesin in the actin-binding sequence.
    J Biol Chem. 1998 Mar 27;273(13):7594-603 PMID: 9516463
  34. Association of the myosin-binding subunit of myosin phosphatase and moesin: dual regulation of moesin phosphorylation by Rho-associated kinase and myosin phosphatase.
    J Cell Biol. 1998 Apr 20;141(2):409-18 PMID: 9548719
  35. Activation of RhoA and SAPK/JNK signalling pathways by the RhoA-specific exchange factor mNET1.
    EMBO J. 1998 Jul 15;17(14):4075-85 PMID: 9670022
  36. Interaction of radixin with Rho small G protein GDP/GTP exchange protein Dbl.
    Oncogene. 1998 Jun 25;16(25):3279-84 PMID: 9681826
  37. Signals from Ras and Rho GTPases interact to regulate expression of p21Waf1/Cip1.
    Nature. 1998 Jul 16;394(6690):295-9 PMID: 9685162
  38. An essential part for Rho-associated kinase in the transcellular invasion of tumor cells.
    Nat Med. 1999 Feb;5(2):221-5 PMID: 9930872
  39. Transformation mediated by RhoA requires activity of ROCK kinases.
    Curr Biol. 1999 Feb 11;9(3):136-45 PMID: 10021386
  40. Regulation of cortical structure by the ezrin-radixin-moesin protein family.
    Curr Opin Cell Biol. 1999 Feb;11(1):109-16 PMID: 10047517
  41. Replacement of threonine 558, a critical site of phosphorylation of moesin in vivo, with aspartate activates F-actin binding of moesin. Regulation by conformational change.
    J Biol Chem. 1999 Apr 30;274(18):12803-10 PMID: 10212266
  42. Merlin: the neurofibromatosis 2 tumor suppressor.
    Biochim Biophys Acta. 1999 Mar 25;1423(2):M29-36 PMID: 10214350
  43. ERM proteins in cell adhesion and membrane dynamics.
    Trends Cell Biol. 1999 May;9(5):187-92 PMID: 10322453
  44. High levels of ezrin expressed by human pancreatic adenocarcinoma cell lines with high metastatic potential.
    Biochem Biophys Res Commun. 1999 May 10;258(2):395-400 PMID: 10329398
  45. Signal-regulated activation of serum response factor is mediated by changes in actin dynamics.
    Cell. 1999 Jul 23;98(2):159-69 PMID: 10428028
  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. Signaling from Rho to the actin cytoskeleton through protein kinases ROCK and LIM-kinase.
    Science. 1999 Aug 6;285(5429):895-8 PMID: 10436159
  48. 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
  49. 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
  50. Essential functions of ezrin in maintenance of cell shape and lamellipodial extension in normal and transformed fibroblasts.
    Curr Biol. 1997 Sep 1;7(9):682-8 PMID: 9285722
  51. 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
  52. Rho GTPases and signaling networks.
    Genes Dev. 1997 Sep 15;11(18):2295-322 PMID: 9308960
  53. Ezrin self-association involves binding of an N-terminal domain to a normally masked C-terminal domain that includes the F-actin binding site.
    Mol Biol Cell. 1995 Aug;6(8):1061-75 PMID: 7579708
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/19.17.4565
Published
2000-09-01
Pages
4565-76
Language
English
Region
England
NLM ID
8208664
PMCID
PMC302055
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/19.17.4565 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: product@genelibs.com

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