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The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1996 May 15;97(10):2289–2298. doi: 10.1172/JCI118671

Defective expression of plectin/HD1 in epidermolysis bullosa simplex with muscular dystrophy.

Y Gache 1, S Chavanas 1, J P Lacour 1, G Wiche 1, K Owaribe 1, G Meneguzzi 1, J P Ortonne 1
PMCID: PMC507309  PMID: 8636409

Abstract

Epidermolysis bullosa simplex with muscular dystrophy (MD-EBS) is a disease characterized by generalized blistering of the skin associated with muscular involvement. We report that the skin of three MD-EBS patients is not reactive with antibodies 6C6, 10F6, or 5B3 raised against the intermediate filament-associated protein plectin. Immunofluorescence and Western analysis of explanted MD-EBS keratinocytes confirmed a deficient expression of plectin, which, in involved skin, correlated with an impaired interaction of the keratin cytoskeleton with the hemidesmosomes. Consistent with lack of reactivity of MD-EBS skin to plectin antibodies, plectin was not detected in skeletal muscles of these patients. Impaired expression of plectin in muscle correlated with an altered labeling pattern of the muscle intermediate filament protein desmin. A deficient immunoreactivity was also observed with the monoclonal antibody HD121 raised against the hemidesmosomal protein HD1. Furthermore, immunofluorescence analysis showed that HD1 is expressed in Z-lines in normal skeletal muscle; whereas this expression is deficient in patient muscle. Colocalization of HD1 and plectin in normal skin and muscle, together with their impaired expression in MD-EBS tissues, strongly suggests that plectin and HD1 are closely related proteins. Our results therefore provide strong evidence that, in MD-EBS patients, the defective expression of plectin results in an aberrant anchorage of cytoskeletal structures in keratinocytes and muscular fibers leading to cell fragility.

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Selected References

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  1. Baudoin C., Miquel C., Blanchet-Bardon C., Gambini C., Meneguzzi G., Ortonne J. P. Herlitz junctional epidermolysis bullosa keratinocytes display heterogeneous defects of nicein/kalinin gene expression. J Clin Invest. 1994 Feb;93(2):862–869. doi: 10.1172/JCI117041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bennett G. S., Fellini S. A., Toyama Y., Holtzer H. Redistribution of intermediate filament subunits during skeletal myogenesis and maturation in vitro. J Cell Biol. 1979 Aug;82(2):577–584. doi: 10.1083/jcb.82.2.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Byers T. J., Kunkel L. M., Watkins S. C. The subcellular distribution of dystrophin in mouse skeletal, cardiac, and smooth muscle. J Cell Biol. 1991 Oct;115(2):411–421. doi: 10.1083/jcb.115.2.411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Campbell K. P. Three muscular dystrophies: loss of cytoskeleton-extracellular matrix linkage. Cell. 1995 Mar 10;80(5):675–679. doi: 10.1016/0092-8674(95)90344-5. [DOI] [PubMed] [Google Scholar]
  5. Chan Y. M., Yu Q. C., LeBlanc-Straceski J., Christiano A., Pulkkinen L., Kucherlapati R. S., Uitto J., Fuchs E. Mutations in the non-helical linker segment L1-2 of keratin 5 in patients with Weber-Cockayne epidermolysis bullosa simplex. J Cell Sci. 1994 Apr;107(Pt 4):765–774. doi: 10.1242/jcs.107.4.765. [DOI] [PubMed] [Google Scholar]
  6. Coulombe P. A., Hutton M. E., Letai A., Hebert A., Paller A. S., Fuchs E. Point mutations in human keratin 14 genes of epidermolysis bullosa simplex patients: genetic and functional analyses. Cell. 1991 Sep 20;66(6):1301–1311. doi: 10.1016/0092-8674(91)90051-y. [DOI] [PubMed] [Google Scholar]
  7. De Luca M., Pellegrini G., Bondanza S., Cremona O., Savoia P., Cancedda R., Marchisio P. C. The control of polarized integrin topography and the organization of adhesion-related cytoskeleton in normal human keratinocytes depend upon number of passages in culture and ionic environment. Exp Cell Res. 1992 Sep;202(1):142–150. doi: 10.1016/0014-4827(92)90413-3. [DOI] [PubMed] [Google Scholar]
  8. Doriguzzi C., Palmucci L., Mongini T., Bertolotto A., Maniscalco M., Chiadò-Piat L., Zina A. M., Bundino S. Congenital muscular dystrophy associated with familial junctional epidermolysis bullosa letalis. Eur Neurol. 1993;33(6):454–460. doi: 10.1159/000116993. [DOI] [PubMed] [Google Scholar]
  9. Ervasti J. M., Campbell K. P. A role for the dystrophin-glycoprotein complex as a transmembrane linker between laminin and actin. J Cell Biol. 1993 Aug;122(4):809–823. doi: 10.1083/jcb.122.4.809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fine J. D., Bauer E. A., Briggaman R. A., Carter D. M., Eady R. A., Esterly N. B., Holbrook K. A., Hurwitz S., Johnson L., Lin A. Revised clinical and laboratory criteria for subtypes of inherited epidermolysis bullosa. A consensus report by the Subcommittee on Diagnosis and Classification of the National Epidermolysis Bullosa Registry. J Am Acad Dermatol. 1991 Jan;24(1):119–135. doi: 10.1016/0190-9622(91)70021-s. [DOI] [PubMed] [Google Scholar]
  11. Fine J. D., Stenn J., Johnson L., Wright T., Bock H. G., Horiguchi Y. Autosomal recessive epidermolysis bullosa simplex. Generalized phenotypic features suggestive of junctional or dystrophic epidermolysis bullosa, and association with neuromuscular diseases. Arch Dermatol. 1989 Jul;125(7):931–938. doi: 10.1001/archderm.125.7.931. [DOI] [PubMed] [Google Scholar]
  12. Foisner R., Feldman B., Sander L., Seifert G., Artlieb U., Wiche G. A panel of monoclonal antibodies to rat plectin: distinction by epitope mapping and immunoreactivity with different tissues and cell lines. Acta Histochem. 1994 Dec;96(4):421–438. doi: 10.1016/S0065-1281(11)80029-2. [DOI] [PubMed] [Google Scholar]
  13. Foisner R., Leichtfried F. E., Herrmann H., Small J. V., Lawson D., Wiche G. Cytoskeleton-associated plectin: in situ localization, in vitro reconstitution, and binding to immobilized intermediate filament proteins. J Cell Biol. 1988 Mar;106(3):723–733. doi: 10.1083/jcb.106.3.723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Guo L., Degenstein L., Dowling J., Yu Q. C., Wollmann R., Perman B., Fuchs E. Gene targeting of BPAG1: abnormalities in mechanical strength and cell migration in stratified epithelia and neurologic degeneration. Cell. 1995 Apr 21;81(2):233–243. doi: 10.1016/0092-8674(95)90333-x. [DOI] [PubMed] [Google Scholar]
  15. Helbling-Leclerc A., Zhang X., Topaloglu H., Cruaud C., Tesson F., Weissenbach J., Tomé F. M., Schwartz K., Fardeau M., Tryggvason K. Mutations in the laminin alpha 2-chain gene (LAMA2) cause merosin-deficient congenital muscular dystrophy. Nat Genet. 1995 Oct;11(2):216–218. doi: 10.1038/ng1095-216. [DOI] [PubMed] [Google Scholar]
  16. Hieda Y., Nishizawa Y., Uematsu J., Owaribe K. Identification of a new hemidesmosomal protein, HD1: a major, high molecular mass component of isolated hemidesmosomes. J Cell Biol. 1992 Mar;116(6):1497–1506. doi: 10.1083/jcb.116.6.1497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hoffman E. P., Kunkel L. M. Dystrophin abnormalities in Duchenne/Becker muscular dystrophy. Neuron. 1989 Jan;2(1):1019–1029. doi: 10.1016/0896-6273(89)90226-2. [DOI] [PubMed] [Google Scholar]
  18. Jones J. C., Asmuth J., Baker S. E., Langhofer M., Roth S. I., Hopkinson S. B. Hemidesmosomes: extracellular matrix/intermediate filament connectors. Exp Cell Res. 1994 Jul;213(1):1–11. doi: 10.1006/excr.1994.1166. [DOI] [PubMed] [Google Scholar]
  19. Kitajima Y., Inoue S., Yaoita H. Abnormal organization of keratin intermediate filaments in cultured keratinocytes of epidermolysis bullosa simplex. Arch Dermatol Res. 1989;281(1):5–10. doi: 10.1007/BF00424265. [DOI] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Lane E. B., Rugg E. L., Navsaria H., Leigh I. M., Heagerty A. H., Ishida-Yamamoto A., Eady R. A. A mutation in the conserved helix termination peptide of keratin 5 in hereditary skin blistering. Nature. 1992 Mar 19;356(6366):244–246. doi: 10.1038/356244a0. [DOI] [PubMed] [Google Scholar]
  22. Matsumura K., Tomé F. M., Collin H., Azibi K., Chaouch M., Kaplan J. C., Fardeau M., Campbell K. P. Deficiency of the 50K dystrophin-associated glycoprotein in severe childhood autosomal recessive muscular dystrophy. Nature. 1992 Sep 24;359(6393):320–322. doi: 10.1038/359320a0. [DOI] [PubMed] [Google Scholar]
  23. Mueller S., Klaus-Kovtun V., Stanley J. R. A 230-kD basic protein is the major bullous pemphigoid antigen. J Invest Dermatol. 1989 Jan;92(1):33–38. doi: 10.1111/1523-1747.ep13070476. [DOI] [PubMed] [Google Scholar]
  24. Niemi K. M., Sommer H., Kero M., Kanerva L., Haltia M. Epidermolysis bullosa simplex associated with muscular dystrophy with recessive inheritance. Arch Dermatol. 1988 Apr;124(4):551–554. [PubMed] [Google Scholar]
  25. Nishizawa Y., Uematsu J., Owaribe K. HD4, a 180 kDa bullous pemphigoid antigen, is a major transmembrane glycoprotein of the hemidesmosome. J Biochem. 1993 Apr;113(4):493–501. doi: 10.1093/oxfordjournals.jbchem.a124072. [DOI] [PubMed] [Google Scholar]
  26. Owaribe K., Nishizawa Y., Franke W. W. Isolation and characterization of hemidesmosomes from bovine corneal epithelial cells. Exp Cell Res. 1991 Feb;192(2):622–630. doi: 10.1016/0014-4827(91)90084-8. [DOI] [PubMed] [Google Scholar]
  27. Rheinwald J. G., Green H. Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell. 1975 Nov;6(3):331–343. doi: 10.1016/s0092-8674(75)80001-8. [DOI] [PubMed] [Google Scholar]
  28. Roberds S. L., Leturcq F., Allamand V., Piccolo F., Jeanpierre M., Anderson R. D., Lim L. E., Lee J. C., Tomé F. M., Romero N. B. Missense mutations in the adhalin gene linked to autosomal recessive muscular dystrophy. Cell. 1994 Aug 26;78(4):625–633. doi: 10.1016/0092-8674(94)90527-4. [DOI] [PubMed] [Google Scholar]
  29. Sarnat H. B. Vimentin and desmin in maturing skeletal muscle and developmental myopathies. Neurology. 1992 Aug;42(8):1616–1624. doi: 10.1212/wnl.42.8.1616. [DOI] [PubMed] [Google Scholar]
  30. Sawamura D., Li K., Chu M. L., Uitto J. Human bullous pemphigoid antigen (BPAG1). Amino acid sequences deduced from cloned cDNAs predict biologically important peptide segments and protein domains. J Biol Chem. 1991 Sep 25;266(27):17784–17790. [PubMed] [Google Scholar]
  31. Simon M., Green H. Enzymatic cross-linking of involucrin and other proteins by keratinocyte particulates in vitro. Cell. 1985 Mar;40(3):677–683. doi: 10.1016/0092-8674(85)90216-8. [DOI] [PubMed] [Google Scholar]
  32. Sonnenberg A., Calafat J., Janssen H., Daams H., van der Raaij-Helmer L. M., Falcioni R., Kennel S. J., Aplin J. D., Baker J., Loizidou M. 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):907–917. doi: 10.1083/jcb.113.4.907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sonnenberg A., Janssen H., Hogervorst F., Calafat J., Hilgers J. A complex of platelet glycoproteins Ic and IIa identified by a rat monoclonal antibody. J Biol Chem. 1987 Jul 25;262(21):10376–10383. [PubMed] [Google Scholar]
  34. Sonnenberg A., Linders C. J., Daams J. H., Kennel S. J. 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):207–217. doi: 10.1242/jcs.96.2.207. [DOI] [PubMed] [Google Scholar]
  35. Spinardi L., Ren Y. L., Sanders R., Giancotti F. G. 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):871–884. doi: 10.1091/mbc.4.9.871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Stepp M. A., Spurr-Michaud S., Tisdale A., Elwell J., Gipson I. K. Alpha 6 beta 4 integrin heterodimer is a component of hemidesmosomes. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8970–8974. doi: 10.1073/pnas.87.22.8970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Stromer M. H. Immunocytochemistry of the muscle cell cytoskeleton. Microsc Res Tech. 1995 Jun 1;31(2):95–105. doi: 10.1002/jemt.1070310202. [DOI] [PubMed] [Google Scholar]
  38. Tanaka T., Korman N. J., Shimizu H., Eady R. A., Klaus-Kovtun V., Cehrs K., Stanley J. R. Production of rabbit antibodies against carboxy-terminal epitopes encoded by bullous pemphigoid cDNA. J Invest Dermatol. 1990 May;94(5):617–623. doi: 10.1111/1523-1747.ep12876200. [DOI] [PubMed] [Google Scholar]
  39. Verrando P., Blanchet-Bardon C., Pisani A., Thomas L., Cambazard F., Eady R. A., Schofield O., Ortonne J. P. Monoclonal antibody GB3 defines a widespread defect of several basement membranes and a keratinocyte dysfunction in patients with lethal junctional epidermolysis bullosa. Lab Invest. 1991 Jan;64(1):85–92. [PubMed] [Google Scholar]
  40. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  41. Wiche G., Becker B., Luber K., Weitzer G., Castañon M. J., Hauptmann R., Stratowa C., Stewart M. 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. doi: 10.1083/jcb.114.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wiche G., Herrmann H., Leichtfried F., Pytela R. Plectin: a high-molecular-weight cytoskeletal polypeptide component that copurifies with intermediate filaments of the vimentin type. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 1):475–482. doi: 10.1101/sqb.1982.046.01.044. [DOI] [PubMed] [Google Scholar]
  43. Wiche G., Krepler R., Artlieb U., Pytela R., Aberer W. Identification of plectin in different human cell types and immunolocalization at epithelial basal cell surface membranes. Exp Cell Res. 1984 Nov;155(1):43–49. doi: 10.1016/0014-4827(84)90766-3. [DOI] [PubMed] [Google Scholar]
  44. Wiche G., Krepler R., Artlieb U., Pytela R., Denk H. Occurrence and immunolocalization of plectin in tissues. J Cell Biol. 1983 Sep;97(3):887–901. doi: 10.1083/jcb.97.3.887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Xu H., Wu X. R., Wewer U. M., Engvall E. Murine muscular dystrophy caused by a mutation in the laminin alpha 2 (Lama2) gene. Nat Genet. 1994 Nov;8(3):297–302. doi: 10.1038/ng1194-297. [DOI] [PubMed] [Google Scholar]

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