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Mol Cell Biol. Oct 1996; 16(10): 5365–5374.
PMCID: PMC231535

Interdependent transcription control elements regulate the expression of the SPRR2A gene during keratinocyte terminal differentiation.

Abstract

Expression of the SPRR2A gene, a member of the small proline-rich family of cornified cell envelope precursor proteins, is strictly linked to keratinocyte terminal differentiation both in vivo and in vitro. In this study, we explored the molecular mechanisms underlying this regulation in transiently transfected primary keratinocytes induced to differentiate in vitro. Deletion mapping and site-directed mutagenesis of SPRR2A promoter-chloramphenicol acetyltransferase constructs indicate that four transcription control elements are essential and sufficient for promoter activity. These elements were further characterized by electrophoretic mobility shift and identified as (i) an inverted octamer doublet, bound by the POU domain factor Oct-11 (Skn-1a/i, Epoc-1), (ii) an interferon-stimulated response element recognized by interferon regulatory factors 1 and 2, (iii) an Ets binding site partially overlapping the interferon-stimulated response element, and (iv) a TG box recognized by the Sp1 family of zinc finger transcription factors. Destruction of a single terminal differentiation element is sufficient to completely abolish transcription from the SPRR2A promoter, indicating that these transcription control elements function in concert in an interdependent manner. Apparently, integration of signals transmitted by the above-mentioned transcription factors is necessary and sufficient to promote gene expression during keratinocyte terminal differentiation.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Adams JC, Watt FM. Fibronectin inhibits the terminal differentiation of human keratinocytes. Nature. 1989 Jul 27;340(6231):307–309. [PubMed]
  • Adams JC, Watt FM. Changes in keratinocyte adhesion during terminal differentiation: reduction in fibronectin binding precedes alpha 5 beta 1 integrin loss from the cell surface. Cell. 1990 Oct 19;63(2):425–435. [PubMed]
  • Andersen B, Schonemann MD, Flynn SE, Pearse RV, 2nd, Singh H, Rosenfeld MG. Skn-1a and Skn-1i: two functionally distinct Oct-2-related factors expressed in epidermis. Science. 1993 Apr 2;260(5104):78–82. [PubMed]
  • Backendorf C, Hohl D. A common origin for cornified envelope proteins? Nat Genet. 1992 Oct;2(2):91–91. [PubMed]
  • Belt PB, Groeneveld H, Teubel WJ, van de Putte P, Backendorf C. Construction and properties of an Epstein-Barr-virus-derived cDNA expression vector for human cells. Gene. 1989 Dec 14;84(2):407–417. [PubMed]
  • Crepieux P, Coll J, Stehelin D. The Ets family of proteins: weak modulators of gene expression in quest for transcriptional partners. Crit Rev Oncog. 1994;5(6):615–638. [PubMed]
  • Fuchs E, Byrne C. The epidermis: rising to the surface. Curr Opin Genet Dev. 1994 Oct;4(5):725–736. [PubMed]
  • Fujimoto W, Marvin KW, George MD, Celli G, Darwiche N, De Luca LM, Jetten AM. Expression of cornifin in squamous differentiating epithelial tissues, including psoriatic and retinoic acid-treated skin. J Invest Dermatol. 1993 Sep;101(3):268–274. [PubMed]
  • Gandarillas A, Watt FM. Changes in expression of members of the fos and jun families and myc network during terminal differentiation of human keratinocytes. Oncogene. 1995 Oct 5;11(7):1403–1407. [PubMed]
  • Gandarillas A, Watt FM. The 5' noncoding region of the mouse involucrin gene: comparison with the human gene and genes encoding other cornified envelope precursors. Mamm Genome. 1995 Sep;6(9):680–682. [PubMed]
  • Garmyn M, Yaar M, Boileau N, Backendorf C, Gilchrest BA. Effect of aging and habitual sun exposure on the genetic response of cultured human keratinocytes to solar-simulated irradiation. J Invest Dermatol. 1992 Dec;99(6):743–748. [PubMed]
  • Gashler A, Sukhatme VP. Early growth response protein 1 (Egr-1): prototype of a zinc-finger family of transcription factors. Prog Nucleic Acid Res Mol Biol. 1995;50:191–224. [PubMed]
  • Gibbs S, Fijneman R, Wiegant J, van Kessel AG, van De Putte P, Backendorf C. Molecular characterization and evolution of the SPRR family of keratinocyte differentiation markers encoding small proline-rich proteins. Genomics. 1993 Jun;16(3):630–637. [PubMed]
  • Gibbs S, Lohman F, Teubel W, van de Putte P, Backendorf C. Characterization of the human spr2 promoter: induction after UV irradiation or TPA treatment and regulation during differentiation of cultured primary keratinocytes. Nucleic Acids Res. 1990 Aug 11;18(15):4401–4407. [PMC free article] [PubMed]
  • Goldsborough AS, Healy LE, Copeland NG, Gilbert DJ, Jenkins NA, Willison KR, Ashworth A. Cloning, chromosomal localization and expression pattern of the POU domain gene Oct-11. Nucleic Acids Res. 1993 Jan 11;21(1):127–134. [PMC free article] [PubMed]
  • Gorman CM, Moffat LF, Howard BH. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. [PMC free article] [PubMed]
  • Greco MA, Lorand L, Lane WS, Baden HP, Parameswaran KN, Kvedar JC. The pancornulins: a group of small proline rich-related cornified envelope precursors with bifunctional capabilities in isopeptide bond formation. J Invest Dermatol. 1995 Feb;104(2):204–210. [PubMed]
  • Green H. Terminal differentiation of cultured human epidermal cells. Cell. 1977 Jun;11(2):405–416. [PubMed]
  • Hagen G, Müller S, Beato M, Suske G. Cloning by recognition site screening of two novel GT box binding proteins: a family of Sp1 related genes. Nucleic Acids Res. 1992 Nov 11;20(21):5519–5525. [PMC free article] [PubMed]
  • Hagen G, Müller S, Beato M, Suske G. Sp1-mediated transcriptional activation is repressed by Sp3. EMBO J. 1994 Aug 15;13(16):3843–3851. [PMC free article] [PubMed]
  • Hamilton TB, Barilla KC, Romaniuk PJ. High affinity binding sites for the Wilms' tumour suppressor protein WT1. Nucleic Acids Res. 1995 Jan 25;23(2):277–284. [PMC free article] [PubMed]
  • Harada H, Fujita T, Miyamoto M, Kimura Y, Maruyama M, Furia A, Miyata T, Taniguchi T. Structurally similar but functionally distinct factors, IRF-1 and IRF-2, bind to the same regulatory elements of IFN and IFN-inducible genes. Cell. 1989 Aug 25;58(4):729–739. [PubMed]
  • Harada H, Kitagawa M, Tanaka N, Yamamoto H, Harada K, Ishihara M, Taniguchi T. Anti-oncogenic and oncogenic potentials of interferon regulatory factors-1 and -2. Science. 1993 Feb 12;259(5097):971–974. [PubMed]
  • Harada H, Willison K, Sakakibara J, Miyamoto M, Fujita T, Taniguchi T. Absence of the type I IFN system in EC cells: transcriptional activator (IRF-1) and repressor (IRF-2) genes are developmentally regulated. Cell. 1990 Oct 19;63(2):303–312. [PubMed]
  • Hardas BD, Zhang J, Trent JM, Elder JT. Direct evidence for homologous sequences on the paracentric regions of human chromosome 1. Genomics. 1994 May 15;21(2):359–363. [PubMed]
  • Hennings H, Michael D, Cheng C, Steinert P, Holbrook K, Yuspa SH. Calcium regulation of growth and differentiation of mouse epidermal cells in culture. Cell. 1980 Jan;19(1):245–254. [PubMed]
  • Herr W, Cleary MA. The POU domain: versatility in transcriptional regulation by a flexible two-in-one DNA-binding domain. Genes Dev. 1995 Jul 15;9(14):1679–1693. [PubMed]
  • Hodivala KJ, Watt FM. Evidence that cadherins play a role in the downregulation of integrin expression that occurs during keratinocyte terminal differentiation. J Cell Biol. 1994 Feb;124(4):589–600. [PMC free article] [PubMed]
  • Hohl D, de Viragh PA, Amiguet-Barras F, Gibbs S, Backendorf C, Huber M. The small proline-rich proteins constitute a multigene family of differentially regulated cornified cell envelope precursor proteins. J Invest Dermatol. 1995 Jun;104(6):902–909. [PubMed]
  • Hohl D, Ruf Olano B, de Viragh PA, Huber M, Detrisac CJ, Schnyder UW, Roop DR. Expression patterns of loricrin in various species and tissues. Differentiation. 1993 Aug;54(1):25–34. [PubMed]
  • Hotchin NA, Gandarillas A, Watt FM. Regulation of cell surface beta 1 integrin levels during keratinocyte terminal differentiation. J Cell Biol. 1995 Mar;128(6):1209–1219. [PMC free article] [PubMed]
  • Hotchin NA, Kovach NL, Watt FM. Functional down-regulation of alpha 5 beta 1 integrin in keratinocytes is reversible but commitment to terminal differentiation is not. J Cell Sci. 1993 Dec;106(Pt 4):1131–1138. [PubMed]
  • Ishida-Yamamoto A, Iizuka H, Manabe M, O'Guin WM, Hohl D, Kartasova T, Kuroki T, Roop DR, Eady RA. Altered distribution of keratinization markers in epidermolytic hyperkeratosis. Arch Dermatol Res. 1995;287(8):705–711. [PubMed]
  • Jensen PK, Bolund L. Low Ca2+ stripping of differentiating cell layers in human epidermal cultures: an in vitro model of epidermal regeneration. Exp Cell Res. 1988 Mar;175(1):63–73. [PubMed]
  • Kadonaga JT, Carner KR, Masiarz FR, Tjian R. Isolation of cDNA encoding transcription factor Sp1 and functional analysis of the DNA binding domain. Cell. 1987 Dec 24;51(6):1079–1090. [PubMed]
  • Kartasova T, van Muijen GN, van Pelt-Heerschap H, van de Putte P. Novel protein in human epidermal keratinocytes: regulation of expression during differentiation. Mol Cell Biol. 1988 May;8(5):2204–2210. [PMC free article] [PubMed]
  • Kartasova T, van de Putte P. Isolation, characterization, and UV-stimulated expression of two families of genes encoding polypeptides of related structure in human epidermal keratinocytes. Mol Cell Biol. 1988 May;8(5):2195–2203. [PMC free article] [PubMed]
  • Koller BH, Orr HT. Cloning and complete sequence of an HLA-A2 gene: analysis of two HLA-A alleles at the nucleotide level. J Immunol. 1985 Apr;134(4):2727–2733. [PubMed]
  • Kunkel TA. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. [PMC free article] [PubMed]
  • Lee JH, Jang SI, Yang JM, Markova NG, Steinert PM. The proximal promoter of the human transglutaminase 3 gene. Stratified squamous epithelial-specific expression in cultured cells is mediated by binding of Sp1 and ets transcription factors to a proximal promoter element. J Biol Chem. 1996 Feb 23;271(8):4561–4568. [PubMed]
  • Martin ME, Piette J, Yaniv M, Tang WJ, Folk WR. Activation of the polyomavirus enhancer by a murine activator protein 1 (AP1) homolog and two contiguous proteins. Proc Natl Acad Sci U S A. 1988 Aug;85(16):5839–5843. [PMC free article] [PubMed]
  • Marvin KW, George MD, Fujimoto W, Saunders NA, Bernacki SH, Jetten AM. Cornifin, a cross-linked envelope precursor in keratinocytes that is down-regulated by retinoids. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):11026–11030. [PMC free article] [PubMed]
  • Matsuyama T, Kimura T, Kitagawa M, Pfeffer K, Kawakami T, Watanabe N, Kündig TM, Amakawa R, Kishihara K, Wakeham A, et al. Targeted disruption of IRF-1 or IRF-2 results in abnormal type I IFN gene induction and aberrant lymphocyte development. Cell. 1993 Oct 8;75(1):83–97. [PubMed]
  • Pellegrini S, Schindler C. Early events in signalling by interferons. Trends Biochem Sci. 1993 Sep;18(9):338–342. [PubMed]
  • Ponec M, Kempenaar JA, De Kloet ER. Corticoids and cultured human epidermal keratinocytes: specific intracellular binding and clinical efficacy. J Invest Dermatol. 1981 Mar;76(3):211–214. [PubMed]
  • Read J, Watt FM. A model for in vitro studies of epidermal homeostasis: proliferation and involucrin synthesis by cultured human keratinocytes during recovery after stripping off the suprabasal layers. J Invest Dermatol. 1988 May;90(5):739–743. [PubMed]
  • Rheinwald JG, 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. [PubMed]
  • Saffer JD, Jackson SP, Annarella MB. Developmental expression of Sp1 in the mouse. Mol Cell Biol. 1991 Apr;11(4):2189–2199. [PMC free article] [PubMed]
  • Saunders NA, Jetten AM. Control of growth regulatory and differentiation-specific genes in human epidermal keratinocytes by interferon gamma. Antagonism by retinoic acid and transforming growth factor beta 1. J Biol Chem. 1994 Jan 21;269(3):2016–2022. [PubMed]
  • Saunders NA, Smith RJ, Jetten AM. Regulation of proliferation-specific and differentiation-specific genes during senescence of human epidermal keratinocyte and mammary epithelial cells. Biochem Biophys Res Commun. 1993 Nov 30;197(1):46–54. [PubMed]
  • Schäfer BW, Wicki R, Engelkamp D, Mattei MG, Heizmann CW. Isolation of a YAC clone covering a cluster of nine S100 genes on human chromosome 1q21: rationale for a new nomenclature of the S100 calcium-binding protein family. Genomics. 1995 Feb 10;25(3):638–643. [PubMed]
  • Schreiber E, Matthias P, Müller MM, Schaffner W. Rapid detection of octamer binding proteins with 'mini-extracts', prepared from a small number of cells. Nucleic Acids Res. 1989 Aug 11;17(15):6419–6419. [PMC free article] [PubMed]
  • Steinert PM, Marekov LN. The proteins elafin, filaggrin, keratin intermediate filaments, loricrin, and small proline-rich proteins 1 and 2 are isodipeptide cross-linked components of the human epidermal cornified cell envelope. J Biol Chem. 1995 Jul 28;270(30):17702–17711. [PubMed]
  • Tamura T, Ishihara M, Lamphier MS, Tanaka N, Oishi I, Aizawa S, Matsuyama T, Mak TW, Taki S, Taniguchi T. An IRF-1-dependent pathway of DNA damage-induced apoptosis in mitogen-activated T lymphocytes. Nature. 1995 Aug 17;376(6541):596–599. [PubMed]
  • Tanaka N, Ishihara M, Kitagawa M, Harada H, Kimura T, Matsuyama T, Lamphier MS, Aizawa S, Mak TW, Taniguchi T. Cellular commitment to oncogene-induced transformation or apoptosis is dependent on the transcription factor IRF-1. Cell. 1994 Jun 17;77(6):829–839. [PubMed]
  • Tanaka N, Taniguchi T. Cytokine gene regulation: regulatory cis-elements and DNA binding factors involved in the interferon system. Adv Immunol. 1992;52:263–281. [PubMed]
  • Volz A, Korge BP, Compton JG, Ziegler A, Steinert PM, Mischke D. Physical mapping of a functional cluster of epidermal differentiation genes on chromosome 1q21. Genomics. 1993 Oct;18(1):92–99. [PubMed]
  • Waskiewicz AJ, Cooper JA. Mitogen and stress response pathways: MAP kinase cascades and phosphatase regulation in mammals and yeast. Curr Opin Cell Biol. 1995 Dec;7(6):798–805. [PubMed]
  • Wasylyk B, Hahn SL, Giovane A. The Ets family of transcription factors. Eur J Biochem. 1993 Jan 15;211(1-2):7–18. [PubMed]
  • Watt FM, Mattey DL, Garrod DR. Calcium-induced reorganization of desmosomal components in cultured human keratinocytes. J Cell Biol. 1984 Dec;99(6):2211–2215. [PMC free article] [PubMed]
  • Westin G, Schaffner W. Heavy metal ions in transcription factors from HeLa cells: Sp1, but not octamer transcription factor requires zinc for DNA binding and for activator function. Nucleic Acids Res. 1988 Jul 11;16(13):5771–5781. [PMC free article] [PubMed]
  • Wirth T, Staudt L, Baltimore D. An octamer oligonucleotide upstream of a TATA motif is sufficient for lymphoid-specific promoter activity. Nature. 1987 Sep 10;329(6135):174–178. [PubMed]
  • Wu RL, Chen TT, Sun TT. Functional importance of an Sp1- and an NFkB-related nuclear protein in a keratinocyte-specific promoter of rabbit K3 keratin gene. J Biol Chem. 1994 Nov 11;269(45):28450–28459. [PubMed]
  • Yaar M, Eller MS, Bhawan J, Harkness DD, DiBenedetto PJ, Gilchrest BA. In vivo and in vitro SPRR1 gene expression in normal and malignant keratinocytes. Exp Cell Res. 1995 Apr;217(2):217–226. [PubMed]
  • Yaar M, Karassik RL, Schnipper LE, Gilchrest BA. Effects of alpha and beta interferons on cultured human keratinocytes. J Invest Dermatol. 1985 Jul;85(1):70–74. [PubMed]
  • Yukawa K, Butz K, Yasui T, Kikutani H, Hoppe-Seyler F. Regulation of human papillomavirus transcription by the differentiation-dependent epithelial factor Epoc-1/skn-1a. J Virol. 1996 Jan;70(1):10–16. [PMC free article] [PubMed]

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