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. 1994 Aug;55(2):289–296.

Recurrent nonsense mutations within the type VII collagen gene in patients with severe recessive dystrophic epidermolysis bullosa.

A Hovnanian 1, L Hilal 1, C Blanchet-Bardon 1, Y de Prost 1, A M Christiano 1, J Uitto 1, M Goossens 1
PMCID: PMC1918351  PMID: 8037207

Abstract

The generalized mutilating form of recessive dystrophic epidermolysis bullosa (i.e., the Hallopeau-Siemens type; HS-RDEB) is a life-threatening disease characterized by extreme mucocutaneous fragility associated with absent or markedly altered anchoring fibrils (AF). Recently, we reported linkage between HS-RDEB and the type VII collagen gene (COL7A1), which encodes the major component of AF. In this study, we investigated 52 unrelated HS-RDEB patients and 2 patients with RDEB inversa for the presence, at CpG dinucleotides, of mutations changing CGA arginine codons to premature stop codons TGA within the COL7A1 gene. Eight exons containing 10 CGA codons located in the amino-terminal domain of the COL7A1 gene were studied. Mutation analysis was performed using denaturing gradient gel electrophoresis of PCR-amplified genomic fragments. Direct sequencing of PCR-amplified products with altered electrophoretic mobility led to the characterization of three premature stop codons, each in a single COL7A1 allele, in four patients. Two patients (one affected with HS-RDEB and the other with RDEB inversa) have the same C-to-T transition at arginine codon 109. Two other HS-RDEB patients have a C-to-T transition at arginine 1213 and 1216, respectively. These nonsense mutations predict the truncation of approximately 56%-92% of the polypeptide, including the collagenous and the noncollagenous NC-2 domains. On the basis of linkage analysis, which showed no evidence for locus heterogeneity in RDEB, it is expected that these patients are compound heterozygotes and have additional mutations on the other COL7A1 allele, leading to impaired AF formation.(ABSTRACT TRUNCATED AT 250 WORDS)

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Selected References

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  1. Amselem S., Sobrier M. L., Duquesnoy P., Rappaport R., Postel-Vinay M. C., Gourmelen M., Dallapiccola B., Goossens M. Recurrent nonsense mutations in the growth hormone receptor from patients with Laron dwarfism. J Clin Invest. 1991 Mar;87(3):1098–1102. doi: 10.1172/JCI115071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baserga S. J., Benz E. J., Jr Nonsense mutations in the human beta-globin gene affect mRNA metabolism. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2056–2060. doi: 10.1073/pnas.85.7.2056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bruckner-Tuderman L., Mitsuhashi Y., Schnyder U. W., Bruckner P. Anchoring fibrils and type VII collagen are absent from skin in severe recessive dystrophic epidermolysis bullosa. J Invest Dermatol. 1989 Jul;93(1):3–9. doi: 10.1111/1523-1747.ep12277331. [DOI] [PubMed] [Google Scholar]
  4. Bruckner-Tuderman L., Rüegger S., Odermatt B., Mitsuhashi Y., Schnyder U. W. Lack of type VII collagen in unaffected skin of patients with severe recessive dystrophic epidermolysis bullosa. Dermatologica. 1988;176(2):57–64. doi: 10.1159/000248673. [DOI] [PubMed] [Google Scholar]
  5. Bruckner-Tuderman L., Winberg J. O., Anton-Lamprecht I., Schnyder U. W., Gedde-Dahl T., Jr Anchoring fibrils, collagen VII, and neutral metalloproteases in recessive dystrophic epidermolysis bullosa inversa. J Invest Dermatol. 1992 Nov;99(5):550–558. doi: 10.1111/1523-1747.ep12667372. [DOI] [PubMed] [Google Scholar]
  6. Burgeson R. E., Lunstrum G. P., Rokosova B., Rimberg C. S., Rosenbaum L. M., Keene D. R. The structure and function of type VII collagen. Ann N Y Acad Sci. 1990;580:32–43. doi: 10.1111/j.1749-6632.1990.tb17915.x. [DOI] [PubMed] [Google Scholar]
  7. Christiano A. M., Anhalt G., Gibbons S., Bauer E. A., Uitto J. Premature termination codons in the type VII collagen gene (COL7A1) underlie severe, mutilating recessive dystrophic epidermolysis bullosa. Genomics. 1994 May 1;21(1):160–168. doi: 10.1006/geno.1994.1238. [DOI] [PubMed] [Google Scholar]
  8. Christiano A. M., Chung-Honet L. C., Hovnanian A., Uitto J. PCR-based detection of two exonic polymorphisms in the human type VII collagen gene (COL7A1) at 3p21.1. Genomics. 1992 Nov;14(3):827–828. doi: 10.1016/s0888-7543(05)80204-6. [DOI] [PubMed] [Google Scholar]
  9. Christiano A. M., Greenspan D. S., Hoffman G. G., Zhang X., Tamai Y., Lin A. N., Dietz H. C., Hovnanian A., Uitto J. A missense mutation in type VII collagen in two affected siblings with recessive dystrophic epidermolysis bullosa. Nat Genet. 1993 May;4(1):62–66. doi: 10.1038/ng0593-62. [DOI] [PubMed] [Google Scholar]
  10. Christiano A. M., Rosenbaum L. M., Chung-Honet L. C., Parente M. G., Woodley D. T., Pan T. C., Zhang R. Z., Chu M. L., Burgeson R. E., Uitto J. The large non-collagenous domain (NC-1) of type VII collagen is amino-terminal and chimeric. Homology to cartilage matrix protein, the type III domains of fibronectin and the A domains of von Willebrand factor. Hum Mol Genet. 1992 Oct;1(7):475–481. doi: 10.1093/hmg/1.7.475. [DOI] [PubMed] [Google Scholar]
  11. Cooper D. N., Youssoufian H. The CpG dinucleotide and human genetic disease. Hum Genet. 1988 Feb;78(2):151–155. doi: 10.1007/BF00278187. [DOI] [PubMed] [Google Scholar]
  12. Dietz H. C., Valle D., Francomano C. A., Kendzior R. J., Jr, Pyeritz R. E., Cutting G. R. The skipping of constitutive exons in vivo induced by nonsense mutations. Science. 1993 Jan 29;259(5095):680–683. doi: 10.1126/science.8430317. [DOI] [PubMed] [Google Scholar]
  13. Epstein E. H., Jr Molecular genetics of epidermolysis bullosa. Science. 1992 May 8;256(5058):799–804. doi: 10.1126/science.1375393. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Gammon W. R., Abernethy M. L., Padilla K. M., Prisayanh P. S., Cook M. E., Wright J., Briggaman R. A., Hunt S. W., 3rd Noncollagenous (NC1) domain of collagen VII resembles multidomain adhesion proteins involved in tissue-specific organization of extracellular matrix. J Invest Dermatol. 1992 Dec;99(6):691–696. doi: 10.1111/1523-1747.ep12614080. [DOI] [PubMed] [Google Scholar]
  16. Greenspan D. S. The carboxyl-terminal half of type VII collagen, including the non-collagenous NC-2 domain and intron/exon organization of the corresponding region of the COL7A1 gene. Hum Mol Genet. 1993 Mar;2(3):273–278. doi: 10.1093/hmg/2.3.273. [DOI] [PubMed] [Google Scholar]
  17. Gruis N. A., Bavinck J. N., Steijlen P. M., van der Schroeff J. G., van Haeringen A., Happle R., Mariman E., van Beersum S. E., Uitto J., Vermeer B. J. Genetic linkage between the collagen VII (COL7A1) gene and the autosomal dominant form of dystrophic epidermolysis bullosa in two Dutch kindreds. J Invest Dermatol. 1992 Nov;99(5):528–530. doi: 10.1111/1523-1747.ep12658066. [DOI] [PubMed] [Google Scholar]
  18. Gyllensten U. B., Erlich H. A. Generation of single-stranded DNA by the polymerase chain reaction and its application to direct sequencing of the HLA-DQA locus. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7652–7656. doi: 10.1073/pnas.85.20.7652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hamosh A., Trapnell B. C., Zeitlin P. L., Montrose-Rafizadeh C., Rosenstein B. J., Crystal R. G., Cutting G. R. Severe deficiency of cystic fibrosis transmembrane conductance regulator messenger RNA carrying nonsense mutations R553X and W1316X in respiratory epithelial cells of patients with cystic fibrosis. J Clin Invest. 1991 Dec;88(6):1880–1885. doi: 10.1172/JCI115510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hashimoto I., Schnyder U. W., Anton-Lamprecht I., Gedde-Dahl T., Jr, Ward S. Ultrastructural studies in epidermolysis bullosa hereditaria. III. Recessive dystrophic types with dermolytic blistering (Hallopeau-Siemens types and inverse type). Arch Dermatol Res. 1976 Aug 27;256(2):137–150. doi: 10.1007/BF00567360. [DOI] [PubMed] [Google Scholar]
  21. Heagerty A. H., Kennedy A. R., Leigh I. M., Purkis P., Eady R. A. Identification of an epidermal basement membrane defect in recessive forms of dystrophic epidermolysis bullosa by LH 7:2 monoclonal antibody: use in diagnosis. Br J Dermatol. 1986 Aug;115(2):125–131. doi: 10.1111/j.1365-2133.1986.tb05707.x. [DOI] [PubMed] [Google Scholar]
  22. Hilal L., Rochat A., Duquesnoy P., Blanchet-Bardon C., Wechsler J., Martin N., Christiano A. M., Barrandon Y., Uitto J., Goossens M. A homozygous insertion-deletion in the type VII collagen gene (COL7A1) in Hallopeau-Siemens dystrophic epidermolysis bullosa. Nat Genet. 1993 Nov;5(3):287–293. doi: 10.1038/ng1193-287. [DOI] [PubMed] [Google Scholar]
  23. Hovnanian A., Duquesnoy P., Blanchet-Bardon C., Knowlton R. G., Amselem S., Lathrop M., Dubertret L., Uitto J., Goossens M. Genetic linkage of recessive dystrophic epidermolysis bullosa to the type VII collagen gene. J Clin Invest. 1992 Sep;90(3):1032–1036. doi: 10.1172/JCI115916. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. John S. W., Weitzner G., Rozen R., Scriver C. R. A rapid procedure for extracting genomic DNA from leukocytes. Nucleic Acids Res. 1991 Jan 25;19(2):408–408. doi: 10.1093/nar/19.2.408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kadowaki T., Kadowaki H., Taylor S. I. A nonsense mutation causing decreased levels of insulin receptor mRNA: detection by a simplified technique for direct sequencing of genomic DNA amplified by the polymerase chain reaction. Proc Natl Acad Sci U S A. 1990 Jan;87(2):658–662. doi: 10.1073/pnas.87.2.658. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Leigh I. M., Eady R. A., Heagerty A. H., Purkis P. E., Whitehead P. A., Burgeson R. E. Type VII collagen is a normal component of epidermal basement membrane, which shows altered expression in recessive dystrophic epidermolysis bullosa. J Invest Dermatol. 1988 May;90(5):639–642. doi: 10.1111/1523-1747.ep12560795. [DOI] [PubMed] [Google Scholar]
  27. Lerman L. S., Silverstein K. Computational simulation of DNA melting and its application to denaturing gradient gel electrophoresis. Methods Enzymol. 1987;155:482–501. doi: 10.1016/0076-6879(87)55032-7. [DOI] [PubMed] [Google Scholar]
  28. Maddalena A., Spence J. E., O'Brien W. E., Nussbaum R. L. Characterization of point mutations in the same arginine codon in three unrelated patients with ornithine transcarbamylase deficiency. J Clin Invest. 1988 Oct;82(4):1353–1358. doi: 10.1172/JCI113738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. McGrath J. A., Ishida-Yamamoto A., O'Grady A., Leigh I. M., Eady R. A. Structural variations in anchoring fibrils in dystrophic epidermolysis bullosa: correlation with type VII collagen expression. J Invest Dermatol. 1993 Apr;100(4):366–372. doi: 10.1111/1523-1747.ep12471830. [DOI] [PubMed] [Google Scholar]
  30. McGrath J. A., Schofield O. M., Mayou B. J., McKee P. H., Eady R. A. Epidermolysis bullosa complicated by squamous cell carcinoma: report of 10 cases. J Cutan Pathol. 1992 Apr;19(2):116–123. doi: 10.1111/j.1600-0560.1992.tb01352.x. [DOI] [PubMed] [Google Scholar]
  31. Myers R. M., Maniatis T., Lerman L. S. Detection and localization of single base changes by denaturing gradient gel electrophoresis. Methods Enzymol. 1987;155:501–527. doi: 10.1016/0076-6879(87)55033-9. [DOI] [PubMed] [Google Scholar]
  32. Parente M. G., Chung L. C., Ryynänen J., Woodley D. T., Wynn K. C., Bauer E. A., Mattei M. G., Chu M. L., Uitto J. Human type VII collagen: cDNA cloning and chromosomal mapping of the gene. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):6931–6935. doi: 10.1073/pnas.88.16.6931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Reiner A. P., Thompson A. R. Screening for nonsense mutations in patients with severe hemophilia A can provide rapid, direct carrier detection. Hum Genet. 1992 Apr;89(1):88–94. doi: 10.1007/BF00207049. [DOI] [PubMed] [Google Scholar]
  34. Ryynänen M., Knowlton R. G., Parente M. G., Chung L. C., Chu M. L., Uitto J. Human type VII collagen: genetic linkage of the gene (COL7A1) on chromosome 3 to dominant dystrophic epidermolysis bullosa. Am J Hum Genet. 1991 Oct;49(4):797–803. [PMC free article] [PubMed] [Google Scholar]
  35. Ryynänen M., Ryynänen J., Sollberg S., Iozzo R. V., Knowlton R. G., Uitto J. Genetic linkage of type VII collagen (COL7A1) to dominant dystrophic epidermolysis bullosa in families with abnormal anchoring fibrils. J Clin Invest. 1992 Mar;89(3):974–980. doi: 10.1172/JCI115680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  37. Sakai L. Y., Keene D. R., Morris N. P., Burgeson R. E. Type VII collagen is a major structural component of anchoring fibrils. J Cell Biol. 1986 Oct;103(4):1577–1586. doi: 10.1083/jcb.103.4.1577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sheffield V. C., Fishman G. A., Beck J. S., Kimura A. E., Stone E. M. Identification of novel rhodopsin mutations associated with retinitis pigmentosa by GC-clamped denaturing gradient gel electrophoresis. Am J Hum Genet. 1991 Oct;49(4):699–706. [PMC free article] [PubMed] [Google Scholar]
  40. Tidman M. J., Eady R. A. Evaluation of anchoring fibrils and other components of the dermal-epidermal junction in dystrophic epidermolysis bullosa by a quantitative ultrastructural technique. J Invest Dermatol. 1985 May;84(5):374–377. doi: 10.1111/1523-1747.ep12265460. [DOI] [PubMed] [Google Scholar]
  41. Youssoufian H., Kazazian H. H., Jr, Phillips D. G., Aronis S., Tsiftis G., Brown V. A., Antonarakis S. E. Recurrent mutations in haemophilia A give evidence for CpG mutation hotspots. 1986 Nov 27-Dec 3Nature. 324(6095):380–382. doi: 10.1038/324380a0. [DOI] [PubMed] [Google Scholar]
  42. al-Imara L., Richards A. J., Eady R. A., Leigh I. M., Farrall M., Pope F. M. Linkage of autosomal dominant dystrophic epidermolysis bullosa in three British families to the marker D3S2 close to the COL7A1 locus. J Med Genet. 1992 Jun;29(6):381–382. doi: 10.1136/jmg.29.6.381. [DOI] [PMC free article] [PubMed] [Google Scholar]

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