Association of Rho-associated Protein Kinase 1 with E-cadherin Complexes is Mediated by P120-catenin

Overview

Journal Mol Biol Cell

Specialties Cell Biology
Molecular Biology

Date 2011 Oct 28

PMID 22031287

Citations 50

Authors

Andrew L Smith

Michael R Dohn

Meredith V Brown

Albert B Reynolds

Affiliations

Soon will be listed here.

Abstract

The dynamic functional linkage of cadherins with the underlying actin cytoskeleton is tightly regulated to achieve proper cell-cell adhesion. p120-catenin (p120) regulates both cadherin stability and actin dynamics, but the relationship between these two functions remains unclear. Using a novel proteomic approach called reversible cross-link immunoprecipitation, or ReCLIP, we previously identified a physical interaction between p120 and Rho-associated protein kinase 1 (ROCK1), a major effector of RhoA. In this paper, we show that a discrete fraction of cellular ROCK1 coimmunoprecipitates with p120 and precisely colocalizes to adherens junctions (AJs). Manipulation of AJs using a calcium-switch assay and cadherin-blocking antibodies indicates direct recruitment of ROCK1 to newly forming junctions. Importantly, we find that p120 links ROCK1 to the cadherin complex, as ROCK1 coimmunoprecipitates with wild-type but not p120-uncoupled E-cadherin. Moreover, depletion of ROCK1 using short-hairpin RNA results in dramatic mislocalization of the cadherin complex and junctional actin. These data are consistent with a model in which p120 dynamically regulates Rho-GTPase activity at the cadherin complex through transient interaction with several of its up- and downstream effectors, including ROCK1.

Citing Articles

Emerging functions of Plakophilin 4 in the control of cell contact dynamics.

Muller L, Hatzfeld M Cell Commun Signal. 2025; 23(1):109.

PMID: 40001215 PMC: 11863852. DOI: 10.1186/s12964-025-02106-1.

Plakophilin 4 controls the spatio-temporal activity of RhoA at adherens junctions to promote cortical actin ring formation and tissue tension.

Muller L, Keil R, Glass M, Hatzfeld M Cell Mol Life Sci. 2024; 81(1):291.

PMID: 38970683 PMC: 11335210. DOI: 10.1007/s00018-024-05329-6.

In mitosis integrins reduce adhesion to extracellular matrix and strengthen adhesion to adjacent cells.

Huber M, Casares-Arias J, Fassler R, Muller D, Strohmeyer N Nat Commun. 2023; 14(1):2143.

PMID: 37059721 PMC: 10104879. DOI: 10.1038/s41467-023-37760-x.

A Novel Variant Identified in a Chinese Family with Blepharocheilodontic Syndrome.

Lin B, Liu Y, Su L, Liu H, Feng H, Yu M Diagnostics (Basel). 2022; 12(12).

PMID: 36552944 PMC: 9777284. DOI: 10.3390/diagnostics12122936.

Nectin4 is a potential therapeutic target for asthma.

Lee P, Choi S, An M, Hwang D, Park S, Baek A Front Immunol. 2022; 13:1049900.

PMID: 36457999 PMC: 9707334. DOI: 10.3389/fimmu.2022.1049900.

References

Birukova A, Zebda N, Cokic I, Fu P, Wu T, Dubrovskyi O . p190RhoGAP mediates protective effects of oxidized phospholipids in the models of ventilator-induced lung injury. Exp Cell Res. 2010; 317(6):859-72. PMC: 3057230. DOI: 10.1016/j.yexcr.2010.11.011. View

Wu J, Mariner D, Thoreson M, Reynolds A . Production and characterization of monoclonal antibodies to the catenin p120ctn. Hybridoma. 1998; 17(2):175-83. DOI: 10.1089/hyb.1998.17.175. View

Zhang J, Betson M, Erasmus J, Zeikos K, Bailly M, Cramer L . Actin at cell-cell junctions is composed of two dynamic and functional populations. J Cell Sci. 2005; 118(Pt 23):5549-62. DOI: 10.1242/jcs.02639. View

Anastasiadis P, Moon S, Thoreson M, Mariner D, Crawford H, Zheng Y . Inhibition of RhoA by p120 catenin. Nat Cell Biol. 2000; 2(9):637-44. DOI: 10.1038/35023588. View

Xia X, Carnahan R, Vaughan M, Wildenberg G, Reynolds A . p120 serine and threonine phosphorylation is controlled by multiple ligand-receptor pathways but not cadherin ligation. Exp Cell Res. 2006; 312(17):3336-48. DOI: 10.1016/j.yexcr.2006.07.007. View

Rimm D, Koslov E, Kebriaei P, Cianci C, Morrow J . Alpha 1(E)-catenin is an actin-binding and -bundling protein mediating the attachment of F-actin to the membrane adhesion complex. Proc Natl Acad Sci U S A. 1995; 92(19):8813-7. PMC: 41057. DOI: 10.1073/pnas.92.19.8813. View

BRAGA V, Betson M, Li X, Lamarche-Vane N . Activation of the small GTPase Rac is sufficient to disrupt cadherin-dependent cell-cell adhesion in normal human keratinocytes. Mol Biol Cell. 2000; 11(11):3703-21. PMC: 15031. DOI: 10.1091/mbc.11.11.3703. View

Castano J, Solanas G, Casagolda D, Raurell I, Villagrasa P, Bustelo X . Specific phosphorylation of p120-catenin regulatory domain differently modulates its binding to RhoA. Mol Cell Biol. 2006; 27(5):1745-57. PMC: 1820477. DOI: 10.1128/MCB.01974-06. View

Molina-Ortiz I, Bartolome R, Hernandez-Varas P, Colo G, Teixido J . Overexpression of E-cadherin on melanoma cells inhibits chemokine-promoted invasion involving p190RhoGAP/p120ctn-dependent inactivation of RhoA. J Biol Chem. 2009; 284(22):15147-57. PMC: 2685696. DOI: 10.1074/jbc.M807834200. View

10.

Hidalgo-Carcedo C, Hooper S, Chaudhry S, Williamson P, Harrington K, Leitinger B . Collective cell migration requires suppression of actomyosin at cell-cell contacts mediated by DDR1 and the cell polarity regulators Par3 and Par6. Nat Cell Biol. 2010; 13(1):49-58. PMC: 3018349. DOI: 10.1038/ncb2133. View

11.

Fukuyama T, Ogita H, Kawakatsu T, Inagaki M, Takai Y . Activation of Rac by cadherin through the c-Src-Rap1-phosphatidylinositol 3-kinase-Vav2 pathway. Oncogene. 2005; 25(1):8-19. DOI: 10.1038/sj.onc.1209010. View

12.

Goodwin M, Kovacs E, Thoreson M, Reynolds A, Yap A . Minimal mutation of the cytoplasmic tail inhibits the ability of E-cadherin to activate Rac but not phosphatidylinositol 3-kinase: direct evidence of a role for cadherin-activated Rac signaling in adhesion and contact formation. J Biol Chem. 2003; 278(23):20533-9. DOI: 10.1074/jbc.M213171200. View

13.

Dohn M, Brown M, Reynolds A . An essential role for p120-catenin in Src- and Rac1-mediated anchorage-independent cell growth. J Cell Biol. 2009; 184(3):437-50. PMC: 2646551. DOI: 10.1083/jcb.200807096. View

14.

Yanagisawa M, Huveldt D, Kreinest P, Lohse C, Cheville J, Parker A . A p120 catenin isoform switch affects Rho activity, induces tumor cell invasion, and predicts metastatic disease. J Biol Chem. 2008; 283(26):18344-54. PMC: 2440599. DOI: 10.1074/jbc.M801192200. View

15.

Ireton R, Davis M, van Hengel J, Mariner D, Barnes K, Thoreson M . A novel role for p120 catenin in E-cadherin function. J Cell Biol. 2002; 159(3):465-76. PMC: 2173073. DOI: 10.1083/jcb.200205115. View

16.

Smith A, Friedman D, Yu H, Carnahan R, Reynolds A . ReCLIP (reversible cross-link immuno-precipitation): an efficient method for interrogation of labile protein complexes. PLoS One. 2011; 6(1):e16206. PMC: 3024417. DOI: 10.1371/journal.pone.0016206. View

17.

Betson M, Lozano E, Zhang J, Braga V . Rac activation upon cell-cell contact formation is dependent on signaling from the epidermal growth factor receptor. J Biol Chem. 2002; 277(40):36962-9. DOI: 10.1074/jbc.M207358200. View

18.

Ishizaki T, Naito M, Fujisawa K, Maekawa M, Watanabe N, Saito Y . p160ROCK, a Rho-associated coiled-coil forming protein kinase, works downstream of Rho and induces focal adhesions. FEBS Lett. 1997; 404(2-3):118-24. DOI: 10.1016/s0014-5793(97)00107-5. View

19.

Shewan A, Maddugoda M, Kraemer A, Stehbens S, Verma S, Kovacs E . Myosin 2 is a key Rho kinase target necessary for the local concentration of E-cadherin at cell-cell contacts. Mol Biol Cell. 2005; 16(10):4531-42. PMC: 1237062. DOI: 10.1091/mbc.e05-04-0330. View

20.

Anderson S, Stone C, Tkach L, SundarRaj N . Rho and Rho-kinase (ROCK) signaling in adherens and gap junction assembly in corneal epithelium. Invest Ophthalmol Vis Sci. 2002; 43(4):978-86. View