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. 1993 Aug;92(2):814–822. doi: 10.1172/JCI116655

The human 230-kD bullous pemphigoid antigen gene (BPAG1). Exon-intron organization and identification of regulatory tissue specific elements in the promoter region.

K Tamai 1, D Sawamura 1, H C Do 1, Y Tamai 1, K Li 1, J Uitto 1
PMCID: PMC294919  PMID: 8349819

Abstract

The 230-kD bullous pemphigoid antigen (BPAG1), a hemidesmosomal protein, is encoded by a gene at the human chromosomal locus 6p11-12. We have elucidated the exon-intron organization of the entire human BPAG1 gene, including approximately 2.6 kb of 5'-flanking DNA. Seven overlapping genomic clones, spanning approximately 20 kb, contained the entire approximately 9 kb coding sequence of BPAG1 and consisted of 22 separate exons, which varied from 78 to 2,810 bp in size. The 5' flanking region of DNA, upstream from the ATG initiation codon for translation, was found to contain several putative transcriptional response elements. Most interestingly, two motifs potentially conferring keratinocyte specific expression to the gene were detected. The presence of such elements was suggested by approximately 20-fold higher expression of a promoter/chloramphenicol acetyl transferase (CAT) construct in normal human epidermal keratinocytes that express the endogenous gene, as compared to several non-expressing cell types. Transient transfections with 5'-deletion clones of the promoter/reporter gene (CAT) constructs identified a region containing a putative tissue specific element, KRE2, which also conferred tissue specificity to the expression of the truncated promoter downstream from this element, however, a mutated derivative of KRE2 was not functional. Detailed knowledge of the structure and regulation of the BPAG1 gene will aid in further elucidation of diseases affecting the cutaneous basement membrane zone.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Angel P., Imagawa M., Chiu R., Stein B., Imbra R. J., Rahmsdorf H. J., Jonat C., Herrlich P., Karin M. Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor. Cell. 1987 Jun 19;49(6):729–739. doi: 10.1016/0092-8674(87)90611-8. [DOI] [PubMed] [Google Scholar]
  2. Beato M. Gene regulation by steroid hormones. Cell. 1989 Feb 10;56(3):335–344. doi: 10.1016/0092-8674(89)90237-7. [DOI] [PubMed] [Google Scholar]
  3. Behr J. P., Demeneix B., Loeffler J. P., Perez-Mutul J. Efficient gene transfer into mammalian primary endocrine cells with lipopolyamine-coated DNA. Proc Natl Acad Sci U S A. 1989 Sep;86(18):6982–6986. doi: 10.1073/pnas.86.18.6982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Birnstiel M. L., Busslinger M., Strub K. Transcription termination and 3' processing: the end is in site! Cell. 1985 Jun;41(2):349–359. doi: 10.1016/s0092-8674(85)80007-6. [DOI] [PubMed] [Google Scholar]
  5. Blessing M., Jorcano J. L., Franke W. W. Enhancer elements directing cell-type-specific expression of cytokeratin genes and changes of the epithelial cytoskeleton by transfections of hybrid cytokeratin genes. EMBO J. 1989 Jan;8(1):117–126. doi: 10.1002/j.1460-2075.1989.tb03355.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Blessing M., Zentgraf H., Jorcano J. L. Differentially expressed bovine cytokeratin genes. Analysis of gene linkage and evolutionary conservation of 5'-upstream sequences. EMBO J. 1987 Mar;6(3):567–575. doi: 10.1002/j.1460-2075.1987.tb04792.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Boyce S. T., Ham R. G. Calcium-regulated differentiation of normal human epidermal keratinocytes in chemically defined clonal culture and serum-free serial culture. J Invest Dermatol. 1983 Jul;81(1 Suppl):33s–40s. doi: 10.1111/1523-1747.ep12540422. [DOI] [PubMed] [Google Scholar]
  8. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  9. Cripe T. P., Haugen T. H., Turk J. P., Tabatabai F., Schmid P. G., 3rd, Dürst M., Gissmann L., Roman A., Turek L. P. Transcriptional regulation of the human papillomavirus-16 E6-E7 promoter by a keratinocyte-dependent enhancer, and by viral E2 trans-activator and repressor gene products: implications for cervical carcinogenesis. EMBO J. 1987 Dec 1;6(12):3745–3753. doi: 10.1002/j.1460-2075.1987.tb02709.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Evans R. M. The steroid and thyroid hormone receptor superfamily. Science. 1988 May 13;240(4854):889–895. doi: 10.1126/science.3283939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fazio M. J., Kähäri V. M., Bashir M. M., Saitta B., Rosenbloom J., Uitto J. Regulation of elastin gene expression: evidence for functional promoter activity in the 5'-flanking region of the human gene. J Invest Dermatol. 1990 Feb;94(2):191–196. doi: 10.1111/1523-1747.ep12874495. [DOI] [PubMed] [Google Scholar]
  12. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Green K. J., Parry D. A., Steinert P. M., Virata M. L., Wagner R. M., Angst B. D., Nilles L. A. Structure of the human desmoplakins. Implications for function in the desmosomal plaque. J Biol Chem. 1990 Feb 15;265(5):2603–2612. [PubMed] [Google Scholar]
  14. Jelinek W. R., Schmid C. W. Repetitive sequences in eukaryotic DNA and their expression. Annu Rev Biochem. 1982;51:813–844. doi: 10.1146/annurev.bi.51.070182.004121. [DOI] [PubMed] [Google Scholar]
  15. Kerr C., Sadowski P. D. Gene 6 exonuclease of bacteriophage T7. II. Mechanism of the reaction. J Biol Chem. 1972 Jan 10;247(1):311–318. [PubMed] [Google Scholar]
  16. Leask A., Byrne C., Fuchs E. Transcription factor AP2 and its role in epidermal-specific gene expression. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):7948–7952. doi: 10.1073/pnas.88.18.7948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Leask A., Rosenberg M., Vassar R., Fuchs E. Regulation of a human epidermal keratin gene: sequences and nuclear factors involved in keratinocyte-specific transcription. Genes Dev. 1990 Nov;4(11):1985–1998. doi: 10.1101/gad.4.11.1985. [DOI] [PubMed] [Google Scholar]
  18. Morrison L. H., Labib R. S., Zone J. J., Diaz L. A., Anhalt G. J. Herpes gestationis autoantibodies recognize a 180-kD human epidermal antigen. J Clin Invest. 1988 Jun;81(6):2023–2026. doi: 10.1172/JCI113554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Mutasim D. F., Takahashi Y., Labib R. S., Anhalt G. J., Patel H. P., Diaz L. A. A pool of bullous pemphigoid antigen(s) is intracellular and associated with the basal cell cytoskeleton-hemidesmosome complex. J Invest Dermatol. 1985 Jan;84(1):47–53. doi: 10.1111/1523-1747.ep12274684. [DOI] [PubMed] [Google Scholar]
  21. Olsen D. R., Hickok N. J., Uitto J. Suppression of ornithine decarboxylase gene expression by retinoids in cultured human keratinocytes. J Invest Dermatol. 1990 Jan;94(1):33–36. doi: 10.1111/1523-1747.ep12873328. [DOI] [PubMed] [Google Scholar]
  22. Padgett R. A., Grabowski P. J., Konarska M. M., Seiler S., Sharp P. A. Splicing of messenger RNA precursors. Annu Rev Biochem. 1986;55:1119–1150. doi: 10.1146/annurev.bi.55.070186.005351. [DOI] [PubMed] [Google Scholar]
  23. Roesler W. J., Vandenbark G. R., Hanson R. W. Cyclic AMP and the induction of eukaryotic gene transcription. J Biol Chem. 1988 Jul 5;263(19):9063–9066. [PubMed] [Google Scholar]
  24. Ryynänen M., Knowlton R. G., Kero M., Sawamura D., Li K. H., Giudice G. J., Diaz L. A., Uitto J. Bullous pemphigoid antigens (BPAGs): identification of RFLPs in human BPAG1 and BPAG2, and exclusion as candidate genes in a large kindred with dominant epidermolysis bullosa simplex. Genomics. 1991 Dec;11(4):1025–1029. doi: 10.1016/0888-7543(91)90028-d. [DOI] [PubMed] [Google Scholar]
  25. Sadler J. R., Waterman M. S., Smith T. F. Regulatory pattern identification in nucleic acid sequences. Nucleic Acids Res. 1983 Apr 11;11(7):2221–2231. doi: 10.1093/nar/11.7.2221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Sawamura D., Li K. H., Nomura K., Sugita Y., Christiano A. M., Uitto J. Bullous pemphigoid antigen: cDNA cloning, cellular expression, and evidence for polymorphism of the human gene. J Invest Dermatol. 1991 Jun;96(6):908–915. doi: 10.1111/1523-1747.ep12475433. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Sawamura D., Nomura K., Sugita Y., Mattei M. G., Chu M. L., Knowlton R., Uitto J. Bullous pemphigoid antigen (BPAG1): cDNA cloning and mapping of the gene to the short arm of human chromosome 6. Genomics. 1990 Dec;8(4):722–726. doi: 10.1016/0888-7543(90)90261-r. [DOI] [PubMed] [Google Scholar]
  30. Snape A. M., Jonas E. A., Sargent T. D. KTF-1, a transcriptional activator of Xenopus embryonic keratin expression. Development. 1990 May;109(1):157–165. doi: 10.1242/dev.109.1.157. [DOI] [PubMed] [Google Scholar]
  31. Snape A. M., Winning R. S., Sargent T. D. Transcription factor AP-2 is tissue-specific in Xenopus and is closely related or identical to keratin transcription factor 1 (KTF-1). Development. 1991 Sep;113(1):283–293. doi: 10.1242/dev.113.1.283. [DOI] [PubMed] [Google Scholar]
  32. Sollberg S., Ryynänen J., Olsen D. R., Uitto J. Transforming growth factor-beta up-regulates the expression of the genes for beta 4 integrin and bullous pemphigoid antigens (BPAG1 and BPAG2) in normal and transformed human keratinocytes. J Invest Dermatol. 1992 Oct;99(4):409–414. doi: 10.1111/1523-1747.ep12616124. [DOI] [PubMed] [Google Scholar]
  33. Stanley J. R., Hawley-Nelson P., Yuspa S. H., Shevach E. M., Katz S. I. Characterization of bullous pemphigoid antigen: a unique basement membrane protein of stratified squamous epithelia. Cell. 1981 Jun;24(3):897–903. doi: 10.1016/0092-8674(81)90115-x. [DOI] [PubMed] [Google Scholar]
  34. Stanley J. R., Tanaka T., Mueller S., Klaus-Kovtun V., Roop D. Isolation of complementary DNA for bullous pemphigoid antigen by use of patients' autoantibodies. J Clin Invest. 1988 Dec;82(6):1864–1870. doi: 10.1172/JCI113803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tanaka T., Parry D. A., Klaus-Kovtun V., Steinert P. M., Stanley J. R. 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):12555–12559. [PubMed] [Google Scholar]
  36. Uitto J., Bauer E. A., Moshell A. N. Symposium on epidermolysis bullosa: molecular biology and pathology of the cutaneous basement membrane zone. Jefferson Medical College, Philadelphia, Pennsylvania, October 4 and 5, 1991. J Invest Dermatol. 1992 Mar;98(3):391–395. doi: 10.1111/1523-1747.ep12499822. [DOI] [PubMed] [Google Scholar]
  37. Ullu E., Tschudi C. Alu sequences are processed 7SL RNA genes. Nature. 1984 Nov 8;312(5990):171–172. doi: 10.1038/312171a0. [DOI] [PubMed] [Google Scholar]
  38. Westgate G. E., Weaver A. C., Couchman J. R. Bullous pemphigoid antigen localization suggests an intracellular association with hemidesmosomes. J Invest Dermatol. 1985 Mar;84(3):218–224. doi: 10.1111/1523-1747.ep12265229. [DOI] [PubMed] [Google Scholar]
  39. Williams T., Tjian R. Analysis of the DNA-binding and activation properties of the human transcription factor AP-2. Genes Dev. 1991 Apr;5(4):670–682. doi: 10.1101/gad.5.4.670. [DOI] [PubMed] [Google Scholar]
  40. de Thé H., Vivanco-Ruiz M. M., Tiollais P., Stunnenberg H., Dejean A. Identification of a retinoic acid responsive element in the retinoic acid receptor beta gene. Nature. 1990 Jan 11;343(6254):177–180. doi: 10.1038/343177a0. [DOI] [PubMed] [Google Scholar]

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