Home LiteratureArticle Details
PMID: 20554761 Published · ppublish English Journal Article Research Support, American Recovery and Reinvestment Act Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Plakophilin 2 couples actomyosin remodeling to desmosomal plaque assembly via RhoA.

Molecular biology of the cell ·Vol. 21 ·No. 16 ·2010-08-15 ·Pages 2844-59

Godsel LM, Dubash AD, Bass-Zubek AE, Amargo EV, Klessner JL, Hobbs RP, Chen X, Green KJ

Abstract

Plakophilin 2 (PKP2), an armadillo family member closely related to p120 catenin (p120ctn), is a constituent of the intercellular adhesive junction, the desmosome. We previously showed that PKP2 loss prevents the incorporation of desmosome precursors enriched in the plaque protein desmoplakin (DP) into newly forming desmosomes, in part by disrupting PKC-dependent regulation of DP assembly competence. On the basis of the observation that DP incorporation into junctions is cytochalasin D-sensitive, here we ask whether PKP2 may also contribute to actin-dependent regulation of desmosome assembly. We demonstrate that PKP2 knockdown impairs cortical actin remodeling after cadherin ligation, without affecting p120ctn expression or localization. Our data suggest that these defects result from the failure of activated RhoA to localize at intercellular interfaces after cell-cell contact and an elevation of cellular RhoA, stress fibers, and other indicators of contractile signaling in squamous cell lines and atrial cardiomyocytes. Consistent with these observations, RhoA activation accelerated DP redistribution to desmosomes during the first hour of junction assembly, whereas sustained RhoA activity compromised desmosome plaque maturation. Together with our previous findings, these data suggest that PKP2 may functionally link RhoA- and PKC-dependent pathways to drive actin reorganization and regulate DP-IF interactions required for normal desmosome assembly.

MeSH Terms
Actins/metabolism Actomyosin/metabolism Animals Cadherins/metabolism Catenins/metabolism Cell Communication Cell Line Cell Line, Tumor Cytoskeleton/metabolism Desmosomes/metabolism Green Fluorescent Proteins/genetics,metabolism Humans Intercellular Junctions/metabolism Microscopy, Fluorescence Myocytes, Cardiac/cytology,metabolism Myosin Light Chains/metabolism Plakophilins/genetics,metabolism Protein Binding Protein Kinase C/metabolism RNA Interference Signal Transduction rhoA GTP-Binding Protein/metabolism
Chemicals
Actins Cadherins Catenins Myosin Light Chains Plakophilins delta catenin Green Fluorescent Proteins Actomyosin Protein Kinase C rhoA GTP-Binding Protein
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Godsel Lisa M
Northwestern University Feinberg School of Medicine, Department of Pathology, Chicago, IL 60611, USA.
Dubash Adi D
Bass-Zubek Amanda E
Amargo Evangeline V
Klessner Jodi L
Hobbs Ryan P
Chen Xinyu
Green Kathleen J
References (73)
73 references, click to expand
  1. p120-catenin and p190RhoGAP regulate cell-cell adhesion by coordinating antagonism between Rac and Rho.
    Cell. 2006 Dec 1;127(5):1027-39 PMID: 17129786
  2. Rho localization in cells and tissues.
    Exp Cell Res. 2004 May 1;295(2):300-14 PMID: 15093731
  3. Plakophilins: multifunctional scaffolds for adhesion and signaling.
    Curr Opin Cell Biol. 2009 Oct;21(5):708-16 PMID: 19674883
  4. Translocation of activated Rho from the cytoplasm to membrane ruffling area, cell-cell adhesion sites and cleavage furrows.
    Oncogene. 1995 Jul 6;11(1):39-48 PMID: 7624130
  5. Desmoplakin II expression is not restricted to stratified epithelia.
    J Cell Sci. 1990 Oct;97 ( Pt 2):247-57 PMID: 2277092
  6. Cell-to-cell adherens junction formation and actin filament organization: similarities and differences between non-polarized fibroblasts and polarized epithelial cells.
    J Cell Sci. 1995 Jan;108 ( Pt 1):127-42 PMID: 7738090
  7. Desmosomes: new perspectives on a classic.
    J Invest Dermatol. 2007 Nov;127(11):2499-515 PMID: 17934502
  8. Plakophilin 2: a critical scaffold for PKC alpha that regulates intercellular junction assembly.
    J Cell Biol. 2008 May 19;181(4):605-13 PMID: 18474624
  9. Cross-talk between adherens junctions and desmosomes depends on plakoglobin.
    J Cell Biol. 1997 Feb 24;136(4):919-34 PMID: 9049256
  10. The function of plakophilin 1 in desmosome assembly and actin filament organization.
    J Cell Biol. 2000 Apr 3;149(1):209-22 PMID: 10747098
  11. Immunofluorescence detection of ezrin/radixin/moesin (ERM) proteins with their carboxyl-terminal threonine phosphorylated in cultured cells and tissues.
    J Cell Sci. 1999 Apr;112 ( Pt 8):1149-58 PMID: 10085250
  12. Directed actin polymerization is the driving force for epithelial cell-cell adhesion.
    Cell. 2000 Jan 21;100(2):209-19 PMID: 10660044
  13. Myosin 2 is a key Rho kinase target necessary for the local concentration of E-cadherin at cell-cell contacts.
    Mol Biol Cell. 2005 Oct;16(10):4531-42 PMID: 16030252
  14. Defining the interactions between intermediate filaments and desmosomes.
    J Cell Biol. 1998 Jun 1;141(5):1229-41 PMID: 9606214
  15. Functional analysis of desmoplakin domains: specification of the interaction with keratin versus vimentin intermediate filament networks.
    J Cell Biol. 1993 Nov;123(3):691-705 PMID: 7693716
  16. Intermediate filament-membrane attachments function synergistically with actin-dependent contacts to regulate intercellular adhesive strength.
    J Cell Biol. 2002 Dec 23;159(6):1005-17 PMID: 12499357
  17. Regulation of Rho GTPases by p120-catenin.
    Curr Opin Cell Biol. 2001 Oct;13(5):604-10 PMID: 11544030
  18. Actin at cell-cell junctions is composed of two dynamic and functional populations.
    J Cell Sci. 2005 Dec 1;118(Pt 23):5549-62 PMID: 16291727
  19. Inhibition of RhoA by p120 catenin.
    Nat Cell Biol. 2000 Sep;2(9):637-44 PMID: 10980705
  20. Actin cable dynamics and Rho/Rock orchestrate a polarized cytoskeletal architecture in the early steps of assembling a stratified epithelium.
    Dev Cell. 2002 Sep;3(3):367-81 PMID: 12361600
  21. Identification of the junctional plaque protein plakophilin 3 in cytoplasmic particles containing RNA-binding proteins and the recruitment of plakophilins 1 and 3 to stress granules.
    Mol Biol Cell. 2006 Mar;17(3):1388-98 PMID: 16407409
  22. Nuclear particles containing RNA polymerase III complexes associated with the junctional plaque protein plakophilin 2.
    Proc Natl Acad Sci U S A. 2001 Jul 3;98(14):7795-800 PMID: 11416169
  23. Cytokinesis: welcome to the Rho zone.
    Trends Cell Biol. 2005 Dec;15(12):651-8 PMID: 16243528
  24. The small GTP-binding protein rho regulates the assembly of focal adhesions and actin stress fibers in response to growth factors.
    Cell. 1992 Aug 7;70(3):389-99 PMID: 1643657
  25. Disorganization of microfilaments and intermediate filaments interferes with the assembly and stability of desmosomes in MDCK epithelial cells.
    Cell Motil Cytoskeleton. 1993;26(2):163-80 PMID: 8287501
  26. Cadherin-directed actin assembly: E-cadherin physically associates with the Arp2/3 complex to direct actin assembly in nascent adhesive contacts.
    Curr Biol. 2002 Mar 5;12(5):379-82 PMID: 11882288
  27. Dissecting temporal and spatial control of cytokinesis with a myosin II Inhibitor.
    Science. 2003 Mar 14;299(5613):1743-7 PMID: 12637748
  28. Skin and heart: une liaison dangereuse.
    Exp Dermatol. 2009 Aug;18(8):658-68 PMID: 19558499
  29. Desmoplakin is essential in epidermal sheet formation.
    Nat Cell Biol. 2001 Dec;3(12):1076-85 PMID: 11781569
  30. Concentric zones of active RhoA and Cdc42 around single cell wounds.
    J Cell Biol. 2005 Jan 31;168(3):429-39 PMID: 15684032
  31. Desmoplakin is required early in development for assembly of desmosomes and cytoskeletal linkage.
    J Cell Biol. 1998 Dec 28;143(7):2009-22 PMID: 9864371
  32. Analysis of activated GAPs and GEFs in cell lysates.
    Methods Enzymol. 2006;406:425-37 PMID: 16472675
  33. Myosin phosphatase: structure, regulation and function.
    Mol Cell Biochem. 2004 Apr;259(1-2):197-209 PMID: 15124925
  34. Making a connection: direct binding between keratin intermediate filaments and desmosomal proteins.
    J Cell Biol. 1994 Nov;127(4):1049-60 PMID: 7525601
  35. Localized zones of Rho and Rac activities drive initiation and expansion of epithelial cell-cell adhesion.
    J Cell Biol. 2007 Jul 30;178(3):517-27 PMID: 17646397
  36. The small GTPases Rho and Rac are required for the establishment of cadherin-dependent cell-cell contacts.
    J Cell Biol. 1997 Jun 16;137(6):1421-31 PMID: 9182672
  37. Plakophilins: Multifunctional proteins or just regulators of desmosomal adhesion?
    Biochim Biophys Acta. 2007 Jan;1773(1):69-77 PMID: 16765467
  38. The relationship between intermediate filaments and microfilaments before and during the formation of desmosomes and adherens-type junctions in mouse epidermal keratinocytes.
    J Cell Biol. 1987 May;104(5):1389-402 PMID: 2437129
  39. Requirement of plakophilin 2 for heart morphogenesis and cardiac junction formation.
    J Cell Biol. 2004 Oct 11;167(1):149-60 PMID: 15479741
  40. Myosin II and mechanotransduction: a balancing act.
    Trends Cell Biol. 2007 Apr;17(4):178-86 PMID: 17320396
  41. A unique and specific interaction between alphaT-catenin and plakophilin-2 in the area composita, the mixed-type junctional structure of cardiac intercalated discs.
    J Cell Sci. 2007 Jun 15;120(Pt 12):2126-36 PMID: 17535849
  42. Mechanism of blebbistatin inhibition of myosin II.
    J Biol Chem. 2004 Aug 20;279(34):35557-63 PMID: 15205456
  43. Defining desmosomal plakophilin-3 interactions.
    J Cell Biol. 2003 Apr 28;161(2):403-16 PMID: 12707304
  44. Role for actin filament turnover and a myosin II motor in cytoskeleton-driven disassembly of the epithelial apical junctional complex.
    Mol Biol Cell. 2004 Jun;15(6):2639-51 PMID: 15047870
  45. Calcium-induced assembly of adherens junctions in keratinocytes.
    J Cell Biol. 1987 Aug;105(2):807-17 PMID: 2442175
  46. p120 catenin regulates the actin cytoskeleton via Rho family GTPases.
    J Cell Biol. 2000 Aug 7;150(3):567-80 PMID: 10931868
  47. Decreased plakophilin-1 expression promotes increased motility in head and neck squamous cell carcinoma cells.
    Cell Commun Adhes. 2007 Mar-Jun;14(2-3):99-109 PMID: 17668353
  48. HL-1 cells: a cardiac muscle cell line that contracts and retains phenotypic characteristics of the adult cardiomyocyte.
    Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):2979-84 PMID: 9501201
  49. Spatiotemporal dynamics of RhoA activity in migrating cells.
    Nature. 2006 Apr 20;440(7087):1069-72 PMID: 16547516
  50. Control of actin polymerization in live and permeabilized fibroblasts.
    J Cell Biol. 1991 Aug;114(3):503-13 PMID: 1860882
  51. Protein binding and functional characterization of plakophilin 2. Evidence for its diverse roles in desmosomes and beta -catenin signaling.
    J Biol Chem. 2002 Mar 22;277(12):10512-22 PMID: 11790773
  52. Desmoplakin assembly dynamics in four dimensions: multiple phases differentially regulated by intermediate filaments and actin.
    J Cell Biol. 2005 Dec 19;171(6):1045-59 PMID: 16365169
  53. An immunofluorescence study of the calcium-induced coordinated reorganization of microfilaments, keratin intermediate filaments, and microtubules in cultured human epidermal keratinocytes.
    J Invest Dermatol. 1991 Dec;97(6):985-94 PMID: 1721081
  54. Interaction of the bullous pemphigoid antigen 1 (BP230) and desmoplakin with intermediate filaments is mediated by distinct sequences within their COOH terminus.
    Mol Biol Cell. 2003 May;14(5):1978-92 PMID: 12802069
  55. Plakophilin 1 interferes with plakoglobin binding to desmoplakin, yet together with plakoglobin promotes clustering of desmosomal plaque complexes at cell-cell borders.
    J Cell Sci. 2001 Feb;114(Pt 4):727-38 PMID: 11171378
  56. Catenins: keeping cells from getting their signals crossed.
    Dev Cell. 2006 Nov;11(5):601-12 PMID: 17084354
  57. Desmosome structure, composition and function.
    Biochim Biophys Acta. 2008 Mar;1778(3):572-87 PMID: 17854763
  58. Differential roles for actin polymerization and a myosin II motor in assembly of the epithelial apical junctional complex.
    Mol Biol Cell. 2005 Jun;16(6):2636-50 PMID: 15800060
  59. Rho GTPase activity zones and transient contractile arrays.
    Bioessays. 2006 Oct;28(10):983-93 PMID: 16998826
  60. Two-hybrid analysis reveals fundamental differences in direct interactions between desmoplakin and cell type-specific intermediate filaments.
    J Biol Chem. 1997 Aug 22;272(34):21495-503 PMID: 9261168
  61. Structures of two intermediate filament-binding fragments of desmoplakin reveal a unique repeat motif structure.
    Nat Struct Biol. 2002 Aug;9(8):612-20 PMID: 12101406
  62. Kazrin regulates keratinocyte cytoskeletal networks, intercellular junctions and differentiation.
    J Cell Sci. 2008 Nov 1;121(Pt 21):3561-9 PMID: 18840647
  63. Cadherin cell-adhesion molecules in human epithelial tissues and carcinomas.
    Cancer Res. 1989 Apr 15;49(8):2128-33 PMID: 2702654
  64. Regulation of the small GTP-binding protein Rho by cell adhesion and the cytoskeleton.
    EMBO J. 1999 Feb 1;18(3):578-85 PMID: 9927417
  65. A novel role for Lsc/p115 RhoGEF and LARG in regulating RhoA activity downstream of adhesion to fibronectin.
    J Cell Sci. 2007 Nov 15;120(Pt 22):3989-98 PMID: 17971419
  66. A role for plakophilin-1 in the initiation of desmosome assembly.
    J Cell Biochem. 2005 Oct 1;96(2):390-403 PMID: 15988759
  67. Biochemical and structural analysis of alpha-catenin in cell-cell contacts.
    Biochem Soc Trans. 2008 Apr;36(Pt 2):141-7 PMID: 18363554
  68. The novel catenin p120cas binds classical cadherins and induces an unusual morphological phenotype in NIH3T3 fibroblasts.
    Exp Cell Res. 1996 Jun 15;225(2):328-37 PMID: 8660921
  69. Carboxyl terminus of Plakophilin-1 recruits it to plasma membrane, whereas amino terminus recruits desmoplakin and promotes desmosome assembly.
    J Biol Chem. 2006 Jun 23;281(25):16962-16970 PMID: 16632867
  70. Protein kinase Calpha-induced p115RhoGEF phosphorylation signals endothelial cytoskeletal rearrangement.
    J Biol Chem. 2003 Aug 1;278(31):28793-8 PMID: 12754211
  71. The area composita of adhering junctions connecting heart muscle cells of vertebrates. VI. Different precursor structures in non-mammalian species.
    Eur J Cell Biol. 2008 Jul;87(7):413-30 PMID: 18420304
  72. Desmosomes at a glance.
    J Cell Sci. 2009 Dec 15;122(Pt 24):4401-7 PMID: 19955337
  73. Plakins: goliaths that link cell junctions and the cytoskeleton.
    Nat Rev Mol Cell Biol. 2004 Jul;5(7):542-53 PMID: 15232572
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1939-4586
Published
2010-08-15
Epub
2010-00-16
Pages
2844-59
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC2921118
Subset
IM
Grants
NIAMS NIH HHS · R01AR43380 · United States
NCI NIH HHS · T32 CA009560 · United States
NIAMS NIH HHS · R01 AR043380 · United States
NIAMS NIH HHS · R01AR041836-17S1 · United States
NIAMS NIH HHS · R01 AR041836 · United States
NIAMS NIH HHS · R01AR41836 · United States
NCI NIH HHS · T32CA009560 · United States
Corrections
ErratumIn
-
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