The functional role of mitogen-activated protein kinase (MAPK) signaling and c-Jun induction in phorbol 12-myristate 13-acetate (PMA)-induced human 12(S)-lipoxygenase gene expression was studied in human epidermoid carcinoma A431 cells. Among the family of MAPK, PMA only increased the activity of extracellular signal-regulated kinase (ERK). Treatment of cells with PD98059, which is an inhibitor of mitogen-activated protein kinase kinase (MEK), decreased the PMA-induced expression of 12(S)-lipoxygenase. Transfection of cells with Ras, Raf and ERK2 dominant negative mutants inhibited the PMA-induced promoter activation of the 12(S)-lipoxygenase gene in all cases. PMA-induced expression of c-Jun was inhibited by pretreatment with PD98059. Following treatment with PMA, the interaction between c-Jun and simian virus 40 promoter factor 1 (Sp1) in cells increased with time. Enhancement of binding between the c-Jun-Sp1 complex and the Sp1 oligonucleotide was observed in cells treated with PMA, suggesting the possible interaction of c-Jun-Sp1 with GC-rich binding sites in the gene promoter. These results indicate that PMA treatment induced ERK activation mainly through the Raf-MEK-ERK signaling pathway following induction of c-Jun expression, and the formation of the c-Jun-Sp1 complex. Finally, PMA activated the promoter activity of the 12(S)-lipoxygenase gene in cells overexpressing protein kinase C (PKC)δ but not PKCα, indicating that PKCδ played the functional role in mediating the gene activation of 12(S)-lipoxygenase induced by PMA.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Biggs JR, Kudlow JE, Kraft AS. The role of the transcription factor Sp1 in regulating the expression of the WAF1/CIP1 gene in U937 leukemic cells. J Biol Chem 271:901–906;1996.
Bruder JT, Heidecker G, Rapp UR. Serum-, TPA-, and Ras-induced expression from AP-1/Ets-driven promoters requires Raf-1 kinase. Genes Dev 6:545–556;1992.
Chang WC, Liu YW, Ning CC, Suzuki H, Yoshimoto T, Yamamoto S. Induction of arachidonate 12-lipoxygenase mRNA by epidermal growth factor in A431 cells. J Biol Chem 268:18734–18739;1993.
Chang WC, Nakao J, Orimo H, Murota S. Effects of reduced glutathione on 12-lipoxygenase pathways in rat platelets. Biochem J 202:771–776;1982.
Chang WC, Ning CC, Lin MT, Huang JD. Epidermal growth factor enhances a microsomal 12-lipoxygenase activity in A431 cells. J Biol Chem 267:3657–3666;1992.
Chen BK, Chang WC. Functional interaction between c-Jun and Sp1 in epidermal growth factor-induced gene expression of human 12(S)-lipoxygenase. Proc Natl Acad Sci USA 97:10406–10411;2000.
Chen BK, Kung HC, Tsai TY, Chang WC. Essential role of mitogen-activated protein kinase pathway and c-Jun induction in epidermal growth factor-induced gene expression of human 12-lipoxygenase. Mol Pharmacol 57:153–161;2000.
Chen BK, Liu YW, Yamamoto S, Chang WC. Overexpression of Ha-ras enhances the transcription of human arachidonate 12-lipoxygenase promoter in A431 cells. Biochim Biophys Acta 1344:270–277;1997.
Chen LC, Chen BK, Liu YW, Chang WC. Induction of 12-lipoxygenase expression by transforming growth factor-alpha in human epidermoid carcinoma A431 cells. FEBS Lett 455:105–110;1999.
Cieslik K, Lee CM, Tang JL, Wu KK. Transcriptional regulation of endothelial nitric-oxide synthase by an interaction between casein kinase 2 and protein phosphatase 2A. J Biol Chem 274:34669–34675;1999.
Daniel S, Zhang S, DePaoli-Roach AA, Kim KH. Dephosphorylation of Sp1 by protein phosphatase 1 is involved in the glucose-mediated activation of the acetyl-CoA carboxylase gene. J Biol Chem 271:14692–14697;1996.
Datto MB, Yu Y, Wang XF. Functional analysis of the transforming growth factor beta responsive elements in the WAF1/Cip1/p21 promoter. J Biol Chem 270:28623–28628;1995.
Derijard B, Raingeaud J, Barrett T, Wu IH, Han J, Ulevitch RJ, Davis RJ. Independent human MAP-kinase signal transduction pathways defined by MEK and MKK isoforms. Science 267:682–685;1995.
Elder JT, Fisher GJ, Lindquist PB, Bennett GL, Pittelkow MR, Coffey JJ, Ellingsworth L, Derynck R, Voorhees JJ. Overexpression of transforming growth factor α in psoriatic epidermis. Science 243:811–814;1989.
Feig LA, Cooper GM. Inhibition of NIH 3T3 cell proliferation by a mutant ras protein with preferential affinity for GDP. Mol Cell Biol 8:3235–3243;1988.
Funk CD, Furci L, FitzGerald GA. Molecular cloning, primary structure, and expression of the human platelet/erythroleukemia cell 12-lipoxygenase. Proc Natl Acad Sci USA 87:5638–5642;1990.
Furstenberger G, Hagedorn H, Jacobi T, Besemfelder E, Stephan M, Lehmann WD, Marks F. Characterization of an 8-lipoxygenase activity induced by the phorbol ester promoter 12-O-tetradecanoyl phorbol-13-acetate in mouse skin in vivo. J Biol Chem 266:15738–15745;1991.
Glasgow WC, Afshari CA, Barrett JC, Eling TE. Modulation of the epidermal growth factor mitogenic response by metabolites of linoleic and arachidonic acid in Syrian hamster embryo fibroblasts. Differential effects in tumor suppressor gene (+) and (−) phenotypes. J Biol Chem 267:10771–10779;1992.
Hamberg M, Samuelsson B. Prostaglandin endoperoxides. Novel transformations of arachidonic acid in human platelets. Proc. Natl Acad Sci USA 71:3400–3404;1974.
Horn F, Marks F, Fisher GJ, Marcelo CL, Voorhees JJ. Decreased protein kinase C activity in psoriatic versus normal epidermis. J Invest Dermatol 88:220–222;1987.
Hussain H, Sornick LP, Shannon VR, Wilson JD, Funk CD, Pentland AP, Holtzman MJ. Epidermis contains platelet-type 12-lipoxygenase that is overexpressed in germinal layer keratinocytes in psoriasis. Am J Physiol 266:C243-C253;1994.
Izumi T, Hoshiko S, Radmark O, Samuelssson B. Cloning of the cDNA for human 12-lipoxygenase. Proc Natl Acad Sci USA 87:7477–7481;1990.
Kardassis D, Papakosta P, Pardali K, Moustakas A. c-Jun transactivates the promoter of the p21WAF1/Cip1 gene by acting as a superactivator of the ubiquitous transcription factor Sp1. J Biol Chem 274:29572–29581;1999.
Khokhlatchev AV, Canagarajah B, Wilsbacher J, Robinson M, Atkinson M, Goldsmith E, Cobb MH. Phosphorylation of the MAP kinase ERK2 promotes its homodimerization and nuclear translocation. Cell 93:605–615;1998.
Leggett RW, Armstrong SA, Barry D, Mueller CR. Sp1 is phosphorylated and its DNA binding activity down-regulated upon terminal differentiation of the liver. J Biol Chem 270:25879–25884;1995.
Liaw YW, Liu YW, Chen BK, Chang WC. Induction of 12-lipoxygenase expression by phorbol 12-myristate 13-acetate in human epidermoid carcinoma A431 cells. Biochim Biophys Acta 1389:23–33;1998.
Liu YW, Arakawa T, Yamamoto S, Chang WC. Transcriptional activation of human 12-lipoxygenase gene promoter is mediated through Sp1 consensus sites in A431 cells. Biochem J 324:133–140;1997.
Liu YW, Asaoka Y, Suzuki H, Yoshimoto T, Yamamoto S, Chang WC. Induction of 12-lipoxygenase expression by epidermal growth factor is mediated by protein kinase C in A431 cells. J Pharmacol Exp Ther 271:567–573;1994.
Moodie SA, Willumsen BM, Weber MJ, Wolfman A. Complexes of Ras. GTP with Raf-1 and mitogen-activated protein kinase kinase. Science 160:1658–1661;1993.
Nishizuka Y. Protein kinase C and lipid signaling for sustained cellular responses. FASEB J 9:484–496;1995.
Ridley AJ, Paterson HF, Johnston CL, Diekmann D, Hall A. The small GTP-binding protein rac regulates growth factor-induced ruffling. Cell 70:401–410;1992.
Robbins DJ, Zhen E, Owaki H, Vanderbilt CA, Ebert D, Geppert TD, Cobb MH. Regulation and properties of extracellular signal-regulated protein kinases 1 and 2 in vitro. J Biol Chem 268:5097–5106;1993.
Shornick LP, Holtzman MJ. A cryptic, microsomal-type arachidonate 12-lipoxygenase is tonically inactivated by oxidation-reduction conditions in cultured epithelial cells. J Biol Chem 268:371–376;1993.
Takahashi Y, Reddy GR, Ueda N, Yamamoto S, Arase S. Arachidonate 12-lipoxygenase of platelet-type in human epidermal cells. J Biol Chem 268:16443–16448;1993.
Ueda Y, Hirai S, Osada S, Suzuki A, Mizuno K, Ohno S. Protein kinase C activates the MEK-ERK pathway in a manner independent of Ras and dependent on Raf. J Biol Chem 271:23512–23519;1996.
About this article
Cite this article
Chen, B., Tsai, T., Huang, H. et al. Functional role of extracellular signal-regulated kinase activation and c-Jun induction in phorbol ester-induced promoter activation of human 12(S)-lipoxygenase gene. J Biomed Sci 9, 156–165 (2002). https://doi.org/10.1007/BF02256027
- Phorbol 12-myristate 13-acetate
- Extracellular signal-regulated kinase