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PMID: 9247637 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Integrin alpha 6 beta 4 forms a complex with the cytoskeletal protein HD1 and induces its redistribution in transfected COS-7 cells.

Molecular biology of the cell ·Vol. 8 ·No. 4 ·1997-04-00 ·Pages 555-66

Niessen CM, Hulsman EH, Rots ES, Sánchez-Aparicio P, Sonnenberg A

Abstract

The integrin alpha 6 beta 4 is a major component of hemidesmosomes, in which it is linked to intermediate filaments. Its presence in these structures is dependent on the beta 4 cytoplasmic domain but it is not known whether beta 4 interacts directly with keratin filaments or by interaction with other proteins. In this study, we have investigated the interaction of GST-cyto beta 4A fusion proteins with cellular proteins and demonstrate that a fragment of beta 4A, consisting of the two pairs of fibronectin type III repeats, separated by the connecting segment, forms a specific complex containing a 500-kDa protein that comigrates with HD1, a hemidesmosomal plaque protein. A similar protein was also bound by a glutathione S-transferase fusion protein containing the cytoplasmic domain of a variant beta 4 subunit (beta 4B), in which a stretch of 53 amino acids is inserted in the connecting segment. Subsequent immunoblot analysis revealed that the 500-kDa protein is in fact HD1. In COS-7 cells, which do not express alpha 6 beta 4 or the hemidesmosomal components BP230 and BP180, HD1 is associated with the cytoskeleton, but after transfecting the cells with cDNAs for human alpha 6 and beta 4, it was, instead, colocalized with alpha 6 beta 4 at the basal side of the cells. The organization of the vimentin, keratin, actin, and tubulin cytoskeletal networks was not affected by the expression of alpha 6 beta 4 in COS-7 cells. The localization of HD1 at the basal side of the cells depends on the same region of beta 4 that forms a complex containing HD1 in vitro, since the expression of alpha 6 with a mutant beta 4 subunit that lacks the four fibronectin type III repeats and the connecting segment did not alter the distribution of HD1. The results indicate that for association of alpha 6 beta 4 with HD1, the cytoplasmic domain of beta 4 is essential. We suggest that this association may be crucial for hemidesmosome assembly.

MeSH Terms
Animals Antigens, Surface/genetics,metabolism Binding Sites Blotting, Western COS Cells/metabolism Cytoplasm/metabolism Cytoskeletal Proteins/chemistry,genetics,metabolism Glutathione Transferase/genetics,metabolism Humans Integrin alpha6beta4 Integrins/genetics,metabolism Intermediate Filament Proteins/genetics,immunology,metabolism Mice Plectin Precipitin Tests Recombinant Fusion Proteins/genetics,metabolism Transfection
Chemicals
Antigens, Surface Cytoskeletal Proteins Integrin alpha6beta4 Integrins Intermediate Filament Proteins PLEC protein, human Plec protein, mouse Plectin Recombinant Fusion Proteins Glutathione Transferase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Niessen C M
Division of Cell Biology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
Hulsman E H
Rots E S
Sánchez-Aparicio P
Sonnenberg A
References (51)
51 references, click to expand
  1. Basement membrane diversity detected by monoclonal antibodies.
    Differentiation. 1984;26(1):49-54 PMID: 6370774
  2. Loss of plectin causes epidermolysis bullosa with muscular dystrophy: cDNA cloning and genomic organization.
    Genes Dev. 1996 Jul 15;10(14):1724-35 PMID: 8698233
  3. Molecular cloning of the CD2 antigen, the T-cell erythrocyte receptor, by a rapid immunoselection procedure.
    Proc Natl Acad Sci U S A. 1987 May;84(10):3365-9 PMID: 2437578
  4. Cytoskeleton-associated plectin: in situ localization, in vitro reconstitution, and binding to immobilized intermediate filament proteins.
    J Cell Biol. 1988 Mar;106(3):723-33 PMID: 3346324
  5. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase.
    Gene. 1988 Jul 15;67(1):31-40 PMID: 3047011
  6. Analysis of the tumor-associated antigen TSP-180. Identity with alpha 6-beta 4 in the integrin superfamily.
    J Biol Chem. 1989 Sep 15;264(26):15515-21 PMID: 2475502
  7. Plectin deficiency results in muscular dystrophy with epidermolysis bullosa.
    Nat Genet. 1996 Aug;13(4):450-7 PMID: 8696340
  8. Deficiency of the integrin beta 4 subunit in junctional epidermolysis bullosa with pyloric atresia: consequences for hemidesmosome formation and adhesion properties.
    J Cell Sci. 1996 Jul;109 ( Pt 7):1695-706 PMID: 8832392
  9. Hemidesmosomes: roles in adhesion, signaling and human diseases.
    Curr Opin Cell Biol. 1996 Oct;8(5):647-56 PMID: 8939649
  10. Characterization of bullous pemphigoid antigen: a unique basement membrane protein of stratified squamous epithelia.
    Cell. 1981 Jun;24(3):897-903 PMID: 7018697
  11. Amino acid sequence of a novel integrin beta 4 subunit and primary expression of the mRNA in epithelial cells.
    EMBO J. 1990 Mar;9(3):757-63 PMID: 2311577
  12. Cloning and sequence analysis of beta-4 cDNA: an integrin subunit that contains a unique 118 kd cytoplasmic domain.
    EMBO J. 1990 Mar;9(3):765-70 PMID: 2311578
  13. Epithelial integrin alpha 6 beta 4: complete primary structure of alpha 6 and variant forms of beta 4.
    J Cell Biol. 1990 Oct;111(4):1593-604 PMID: 1976638
  14. The alpha 6 beta 1 (VLA-6) and alpha 6 beta 4 protein complexes: tissue distribution and biochemical properties.
    J Cell Sci. 1990 Jun;96 ( Pt 2):207-17 PMID: 1698797
  15. Alpha 6 beta 4 integrin heterodimer is a component of hemidesmosomes.
    Proc Natl Acad Sci U S A. 1990 Nov;87(22):8970-4 PMID: 2247472
  16. Isolation and characterization of hemidesmosomes from bovine corneal epithelial cells.
    Exp Cell Res. 1991 Feb;192(2):622-30 PMID: 1988297
  17. A novel hemidesmosomal plaque component: tissue distribution and incorporation into assembling hemidesmosomes in an in vitro model.
    Exp Cell Res. 1991 May;194(1):139-46 PMID: 2015847
  18. Integrin alpha 6/beta 4 complex is located in hemidesmosomes, suggesting a major role in epidermal cell-basement membrane adhesion.
    J Cell Biol. 1991 May;113(4):907-17 PMID: 2026654
  19. Cloning and sequencing of rat plectin indicates a 466-kD polypeptide chain with a three-domain structure based on a central alpha-helical coiled coil.
    J Cell Biol. 1991 Jul;114(1):83-99 PMID: 2050743
  20. Comparison of molecularly cloned bullous pemphigoid antigen to desmoplakin I confirms that they define a new family of cell adhesion junction plaque proteins.
    J Biol Chem. 1991 Jul 5;266(19):12555-9 PMID: 1712022
  21. Intermediate filament-associated proteins.
    Curr Opin Cell Biol. 1991 Feb;3(1):75-81 PMID: 1854487
  22. A function for the integrin alpha 6 beta 4 in the hemidesmosome.
    Cell Regul. 1991 Jun;2(6):427-38 PMID: 1883873
  23. Surface relocation of alpha 6 beta 4 integrins and assembly of hemidesmosomes in an in vitro model of wound healing.
    J Cell Biol. 1991 Dec;115(6):1737-50 PMID: 1757471
  24. Identification of a new hemidesmosomal protein, HD1: a major, high molecular mass component of isolated hemidesmosomes.
    J Cell Biol. 1992 Mar;116(6):1497-506 PMID: 1541639
  25. Integrins: versatility, modulation, and signaling in cell adhesion.
    Cell. 1992 Apr 3;69(1):11-25 PMID: 1555235
  26. The beta 4 subunit of the integrin family is displayed on a restricted subset of endothelium in mice.
    J Cell Sci. 1992 Jan;101 ( Pt 1):145-50 PMID: 1569120
  27. Developmentally regulated expression of the beta 4 integrin on immature mouse thymocytes.
    J Immunol. 1992 Jul 15;149(2):421-8 PMID: 1385605
  28. Proteolytic processing of endogenous and recombinant beta 4 integrin subunit.
    J Cell Biol. 1992 Aug;118(4):951-9 PMID: 1500432
  29. Comparative structural analysis of desmoplakin, bullous pemphigoid antigen and plectin: members of a new gene family involved in organization of intermediate filaments.
    Int J Biol Macromol. 1992 Jun;14(3):145-53 PMID: 1390446
  30. Biochemical characterization and tissue distribution of the A and B variants of the integrin alpha 6 subunit.
    J Cell Biol. 1993 Apr;121(1):179-91 PMID: 7681434
  31. Expression of plectin mutant cDNA in cultured cells indicates a role of COOH-terminal domain in intermediate filament association.
    J Cell Biol. 1993 May;121(3):607-19 PMID: 8486740
  32. The beta 4 subunit cytoplasmic domain mediates the interaction of alpha 6 beta 4 integrin with the cytoskeleton of hemidesmosomes.
    Mol Biol Cell. 1993 Sep;4(9):871-84 PMID: 8257791
  33. Formation of hemidesmosomes in cells of a transformed murine mammary tumor cell line and mechanisms involved in adherence of these cells to laminin and kalinin.
    J Cell Sci. 1993 Dec;106 ( Pt 4):1083-102 PMID: 8126093
  34. IFAP 300 is common to desmosomes and hemidesmosomes and is a possible linker of intermediate filaments to these junctions.
    J Cell Biol. 1994 Apr;125(1):159-70 PMID: 8138568
  35. The alpha 6 beta 4 integrin is a receptor for both laminin and kalinin.
    Exp Cell Res. 1994 Apr;211(2):360-7 PMID: 8143784
  36. Proteolytic cleavage of the integrin beta 4 subunit.
    Exp Cell Res. 1994 May;212(1):2-9 PMID: 8174638
  37. Expression of the integrin alpha 6 beta 4 in peripheral nerves: localization in Schwann and perineural cells and different variants of the beta 4 subunit.
    J Cell Sci. 1994 Feb;107 ( Pt 2):543-52 PMID: 8207077
  38. Demonstration of type II hemidesmosomes in a mammary gland epithelial cell line, BMGE-H.
    J Biochem. 1994 Mar;115(3):469-76 PMID: 8056759
  39. A novel structural variant of the human beta 4 integrin cDNA.
    Cell Adhes Commun. 1994 Apr;2(1):1-6 PMID: 7982032
  40. The mucin epiglycanin on TA3/Ha carcinoma cells prevents alpha 6 beta 4-mediated adhesion to laminin and kalinin and E-cadherin-mediated cell-cell interaction.
    J Cell Biol. 1994 Dec;127(6 Pt 2):2071-80 PMID: 7528749
  41. 180-kD bullous pemphigoid antigen (BP180) is deficient in generalized atrophic benign epidermolysis bullosa.
    J Clin Invest. 1995 Mar;95(3):1345-52 PMID: 7883981
  42. Gene targeting of BPAG1: abnormalities in mechanical strength and cell migration in stratified epithelia and neurologic degeneration.
    Cell. 1995 Apr 21;81(2):233-43 PMID: 7736575
  43. Molecular genetic studies of a human epidermal autoantigen (the 180-kD bullous pemphigoid antigen/BP180): identification of functionally important sequences within the BP180 molecule and evidence for an interaction between BP180 and alpha 6 integrin.
    J Cell Biol. 1995 Jul;130(1):117-25 PMID: 7790367
  44. Signal transduction by the alpha 6 beta 4 integrin: distinct beta 4 subunit sites mediate recruitment of Shc/Grb2 and association with the cytoskeleton of hemidesmosomes.
    EMBO J. 1995 Sep 15;14(18):4470-81 PMID: 7556090
  45. Mutations in the 180-kD bullous pemphigoid antigen (BPAG2), a hemidesmosomal transmembrane collagen (COL17A1), in generalized atrophic benign epidermolysis bullosa.
    Nat Genet. 1995 Sep;11(1):83-6 PMID: 7550320
  46. Defective expression of plectin/HD1 in epidermolysis bullosa simplex with muscular dystrophy.
    J Clin Invest. 1996 May 15;97(10):2289-98 PMID: 8636409
  47. Desmosomes and hemidesmosomes: structure and function of molecular components.
    FASEB J. 1996 Jun;10(8):871-81 PMID: 8666164
  48. Epithelial detachment due to absence of hemidesmosomes in integrin beta 4 null mice.
    Nat Genet. 1996 Jul;13(3):366-9 PMID: 8673140
  49. Absence of integrin alpha 6 leads to epidermolysis bullosa and neonatal death in mice.
    Nat Genet. 1996 Jul;13(3):370-3 PMID: 8673141
  50. Beta4 integrin is required for hemidesmosome formation, cell adhesion and cell survival.
    J Cell Biol. 1996 Jul;134(2):559-72 PMID: 8707838
  51. Identification of plectin in different human cell types and immunolocalization at epithelial basal cell surface membranes.
    Exp Cell Res. 1984 Nov;155(1):43-9 PMID: 6386498
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.8.4.555
Published
1997-04-00
Pages
555-66
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC276108
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.8.4.555 PMC Full Text View journal details
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