Abstract
Involucrin is one of the precursor proteins of the cornified cell envelope of keratinocytes, and is expressed during the later stages of keratinocyte differentiation. Interferon-gamma (IFN-gamma), a pleiotropic cytokine with anti-proliferative and immunomodulatory activities, is also a potent inducer of squamous differentiation. Using cultured normal human keratinocytes (NHK cells) and simian virus 40-transformed human keratinocytes (SVHK cells), we investigated the effects of IFN-gamma on involucrin gene expression. Expression of involucrin was increased by about 3-fold after treating NHK cells with IFN-gamma (100 units/ml). Northern blot analyses revealed that IFN-gamma increased the expression of involucrin mRNA. The fragment +42 to -2463 in the 5'-flanking region of the human involucrin gene was subcloned into a luciferase reporter vector and the construct (p2463Luc) was transfected into SVHK cells. p2463Luc produced a 3-fold increase in luciferase activity after IFN-gamma treatment. Sequence analysis detected two putative IFN-gamma-responsive regions [G1 (positions -883 to -874) and G2 (-784 to -775)]. Deletion analyses of the p2463Luc vector revealed that the G1 region is critical for the IFN-gamma-dependent up-regulation of the involucrin gene. Gel-shift analyses revealed that STAT1 (signal transduction and activators of transcription 1) protein bound to the G1 region and that involucrin promoter activity was augmented by transfection of a STAT1 expression vector in the presence of IFN-gamma. In contrast, transfection of a STAT1 dominant-negative expression vector suppressed the IFN-gamma-dependent up-regulation of involucrin promoter activity. These results indicate that IFN-gamma stimulates expression of the human involucrin gene via the G1 (-883 to -874) region of the involucrin gene promoter.
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- Bach E. A., Aguet M., Schreiber R. D. The IFN gamma receptor: a paradigm for cytokine receptor signaling. Annu Rev Immunol. 1997;15:563–591. doi: 10.1146/annurev.immunol.15.1.563. [DOI] [PubMed] [Google Scholar]
- Banks-Schlegel S., Green H. Involucrin synthesis and tissue assembly by keratinocytes in natural and cultured human epithelia. J Cell Biol. 1981 Sep;90(3):732–737. doi: 10.1083/jcb.90.3.732. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barker J. N., Karabin G. D., Stoof T. J., Sarma V. J., Dixit V. M., Nickoloff B. J. Detection of interferon-gamma mRNA in psoriatic epidermis by polymerase chain reaction. J Dermatol Sci. 1991 Mar;2(2):106–111. doi: 10.1016/0923-1811(91)90019-t. [DOI] [PubMed] [Google Scholar]
- Brysk M. M., Bell T., Hoida C., Tyring S. K., Rajaraman S. Interferon-gamma modulates terminal differentiation and the expression of desquamin in cultured keratinocytes. Exp Cell Res. 1991 Dec;197(2):140–147. doi: 10.1016/0014-4827(91)90415-q. [DOI] [PubMed] [Google Scholar]
- Crish J. F., Zaim T. M., Eckert R. L. The distal regulatory region of the human involucrin promoter is required for expression in epidermis. J Biol Chem. 1998 Nov 13;273(46):30460–30465. doi: 10.1074/jbc.273.46.30460. [DOI] [PubMed] [Google Scholar]
- Darnell J. E., Jr, Kerr I. M., Stark G. R. Jak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins. Science. 1994 Jun 3;264(5164):1415–1421. doi: 10.1126/science.8197455. [DOI] [PubMed] [Google Scholar]
- Dover R., Watt F. M. Measurement of the rate of epidermal terminal differentiation: expression of involucrin by S-phase keratinocytes in culture and in psoriatic plaques. J Invest Dermatol. 1987 Oct;89(4):349–352. doi: 10.1111/1523-1747.ep12471751. [DOI] [PubMed] [Google Scholar]
- Eckert R. L., Green H. Structure and evolution of the human involucrin gene. Cell. 1986 Aug 15;46(4):583–589. doi: 10.1016/0092-8674(86)90884-6. [DOI] [PubMed] [Google Scholar]
- Eckert R. L., Yaffe M. B., Crish J. F., Murthy S., Rorke E. A., Welter J. F. Involucrin--structure and role in envelope assembly. J Invest Dermatol. 1993 May;100(5):613–617. doi: 10.1111/1523-1747.ep12472288. [DOI] [PubMed] [Google Scholar]
- Eckert R. L., Yaffe M. B., Crish J. F., Murthy S., Rorke E. A., Welter J. F. Involucrin--structure and role in envelope assembly. J Invest Dermatol. 1993 May;100(5):613–617. doi: 10.1111/1523-1747.ep12472288. [DOI] [PubMed] [Google Scholar]
- Efimova T., LaCelle P., Welter J. F., Eckert R. L. Regulation of human involucrin promoter activity by a protein kinase C, Ras, MEKK1, MEK3, p38/RK, AP1 signal transduction pathway. J Biol Chem. 1998 Sep 18;273(38):24387–24395. doi: 10.1074/jbc.273.38.24387. [DOI] [PubMed] [Google Scholar]
- Etoh Y., Simon M., Green H. Involucrin acts as a transglutaminase substrate at multiple sites. Biochem Biophys Res Commun. 1986 Apr 14;136(1):51–56. doi: 10.1016/0006-291x(86)90875-2. [DOI] [PubMed] [Google Scholar]
- Felgner P. L., Gadek T. R., Holm M., Roman R., Chan H. W., Wenz M., Northrop J. P., Ringold G. M., Danielsen M. Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7413–7417. doi: 10.1073/pnas.84.21.7413. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L. R. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene. 1989 Apr 15;77(1):51–59. doi: 10.1016/0378-1119(89)90358-2. [DOI] [PubMed] [Google Scholar]
- Hohl D. Cornified cell envelope. Dermatologica. 1990;180(4):201–211. doi: 10.1159/000248031. [DOI] [PubMed] [Google Scholar]
- Hohl D., Mehrel T., Lichti U., Turner M. L., Roop D. R., Steinert P. M. Characterization of human loricrin. Structure and function of a new class of epidermal cell envelope proteins. J Biol Chem. 1991 Apr 5;266(10):6626–6636. [PubMed] [Google Scholar]
- Iizuka H., Honda H., Ishida-Yamamoto A. Epidermal remodeling in psoriasis (II): a quantitative analysis of the epidermal architecture. J Invest Dermatol. 1997 Dec;109(6):806–810. doi: 10.1111/1523-1747.ep12341002. [DOI] [PubMed] [Google Scholar]
- Ishida-Yamamoto A., Kartasova T., Matsuo S., Kuroki T., Iizuka H. Involucrin and SPRR are synthesized sequentially in differentiating cultured epidermal cells. J Invest Dermatol. 1997 Jan;108(1):12–16. doi: 10.1111/1523-1747.ep12285611. [DOI] [PubMed] [Google Scholar]
- 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. doi: 10.1128/mcb.8.5.2195. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kashihara-Sawami M., Norris D. A. The state of differentiation of cultured human keratinocytes determines the level of intercellular adhesion molecule-1 (ICAM-1) expression induced by gamma interferon. J Invest Dermatol. 1992 May;98(5):741–747. doi: 10.1111/1523-1747.ep12499938. [DOI] [PubMed] [Google Scholar]
- Lopez-Bayghen E., Vega A., Cadena A., Granados S. E., Jave L. F., Gariglio P., Alvarez-Salas L. M. Transcriptional analysis of the 5'-noncoding region of the human involucrin gene. J Biol Chem. 1996 Jan 5;271(1):512–520. doi: 10.1074/jbc.271.1.512. [DOI] [PubMed] [Google Scholar]
- Naik S. M., Shibagaki N., Li L. J., Quinlan K. L., Paxton L. L., Caughman S. W. Interferon gamma-dependent induction of human intercellular adhesion molecule-1 gene expression involves activation of a distinct STAT protein complex. J Biol Chem. 1997 Jan 10;272(2):1283–1290. doi: 10.1074/jbc.272.2.1283. [DOI] [PubMed] [Google Scholar]
- Nakajima K., Yamanaka Y., Nakae K., Kojima H., Ichiba M., Kiuchi N., Kitaoka T., Fukada T., Hibi M., Hirano T. A central role for Stat3 in IL-6-induced regulation of growth and differentiation in M1 leukemia cells. EMBO J. 1996 Jul 15;15(14):3651–3658. [PMC free article] [PubMed] [Google Scholar]
- Nickoloff B. J., Mahrle G., Morhenn V. Ultrastructural effects of recombinant gamma-interferon on cultured human keratinocytes. Ultrastruct Pathol. 1986;10(1):17–21. doi: 10.3109/01913128609015559. [DOI] [PubMed] [Google Scholar]
- Ruhrberg C., Hajibagheri M. A., Simon M., Dooley T. P., Watt F. M. Envoplakin, a novel precursor of the cornified envelope that has homology to desmoplakin. J Cell Biol. 1996 Aug;134(3):715–729. doi: 10.1083/jcb.134.3.715. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Saunders N. A., Jetten A. M. 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] [Google Scholar]
- Scheynius A., Fransson J., Johansson C., Hammar H., Baker B., Fry L., Valdimarsson H. Expression of interferon-gamma receptors in normal and psoriatic skin. J Invest Dermatol. 1992 Feb;98(2):255–258. doi: 10.1111/1523-1747.ep12556086. [DOI] [PubMed] [Google Scholar]
- Stark G. R., Kerr I. M., Williams B. R., Silverman R. H., Schreiber R. D. How cells respond to interferons. Annu Rev Biochem. 1998;67:227–264. doi: 10.1146/annurev.biochem.67.1.227. [DOI] [PubMed] [Google Scholar]
- Steinberg M. L., Defendi V. Transformation and immortalization of human keratinocytes by SV40. J Invest Dermatol. 1983 Jul;81(1 Suppl):131s–136s. doi: 10.1111/1523-1747.ep12540905. [DOI] [PubMed] [Google Scholar]
- Steinert P. M., Marekov L. N. 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. doi: 10.1074/jbc.270.30.17702. [DOI] [PubMed] [Google Scholar]
- Takahashi H., Asano K., Kinouchi M., Ishida-Yamamoto A., Wuepper K. D., Iizuka H. Structure and transcriptional regulation of the human cystatin A gene. The 12-O-tetradecanoylphorbol-13-acetate (TPA) responsive element-2 site (-272 to -278) on cystatin A gene is critical for TPA-dependent regulation. J Biol Chem. 1998 Jul 10;273(28):17375–17380. doi: 10.1074/jbc.273.28.17375. [DOI] [PubMed] [Google Scholar]
- Takahashi H., Iizuka H. Analysis of the 5'-upstream promoter region of human involucrin gene: activation by 12-O-tetradecanoylphorbol-13-acetate. J Invest Dermatol. 1993 Jan;100(1):10–15. doi: 10.1111/1523-1747.ep12349867. [DOI] [PubMed] [Google Scholar]
- Takahashi H., Kobayashi H., Matsuo S., Iizuka H. Repression of involucrin gene expression by transcriptional enhancer factor 1 (TEF-1). Arch Dermatol Res. 1995;287(8):740–746. doi: 10.1007/BF01105799. [DOI] [PubMed] [Google Scholar]
- Takematsu H., Tagami H. Interleukin 2, soluble interleukin 2 receptor, and interferon-gamma in the suction blister fluids from psoriatic skin. Arch Dermatol Res. 1990;282(3):149–152. doi: 10.1007/BF00372613. [DOI] [PubMed] [Google Scholar]
- Ulloa L., Doody J., Massagué J. Inhibition of transforming growth factor-beta/SMAD signalling by the interferon-gamma/STAT pathway. Nature. 1999 Feb 25;397(6721):710–713. doi: 10.1038/17826. [DOI] [PubMed] [Google Scholar]
- Uyemura K., Yamamura M., Fivenson D. F., Modlin R. L., Nickoloff B. J. The cytokine network in lesional and lesion-free psoriatic skin is characterized by a T-helper type 1 cell-mediated response. J Invest Dermatol. 1993 Nov;101(5):701–705. doi: 10.1111/1523-1747.ep12371679. [DOI] [PubMed] [Google Scholar]
- Vollmer S., Menssen A., Trommler P., Schendel D., Prinz J. C. T lymphocytes derived from skin lesions of patients with psoriasis vulgaris express a novel cytokine pattern that is distinct from that of T helper type 1 and T helper type 2 cells. Eur J Immunol. 1994 Oct;24(10):2377–2382. doi: 10.1002/eji.1830241018. [DOI] [PubMed] [Google Scholar]
- Welter J. F., Crish J. F., Agarwal C., Eckert R. L. Fos-related antigen (Fra-1), junB, and junD activate human involucrin promoter transcription by binding to proximal and distal AP1 sites to mediate phorbol ester effects on promoter activity. J Biol Chem. 1995 May 26;270(21):12614–12622. doi: 10.1074/jbc.270.21.12614. [DOI] [PubMed] [Google Scholar]
- Welter J. F., Gali H., Crish J. F., Eckert R. L. Regulation of human involucrin promoter activity by POU domain proteins. J Biol Chem. 1996 Jun 21;271(25):14727–14733. doi: 10.1074/jbc.271.25.14727. [DOI] [PubMed] [Google Scholar]
- van den Oord J. J., De Ley M., De Wolf-Peeters C. Distribution of interferon-gamma receptors in normal and psoriatic skin. Pathol Res Pract. 1995 Jul;191(6):530–534. doi: 10.1016/S0344-0338(11)80872-1. [DOI] [PubMed] [Google Scholar]