Abstract
The induction of the proenkephalin gene by nicotine has been characterized in bovine adrenal medullary chromaffin cells. Nicotine (10 μM) caused an approximately fourfold increase in the proenkephalin mRNA levels within 24 h. The half-life of the proenkephalin mRNA in nicotine-stimulated cells was similar to that in control cells (about 13 h), indicating that nicotine does not affect mRNA stability but acts at the levels of proenkephalin gene transcription. This was also supported by experiments showing that the expression of a proenkephalin chloramphenicol acetyl transferase reporter gene (PENKCAT-153/+50) containing 153 nucleotides of upstream promoter sequences is increased (about twofold) by nicotine after transient transfection in the chromaffin cells. In addition, nicotine induced a marked elevation of the immediate early gene mRNAs c-fos, c-jun, and jun-B. Maximally increased levels for c-fos mRNA (about 100-fold) were obtained after 20 min. c-jun and jun-B were increased three- to fivefold 60 min after nicotine addition. The expression of PENKCAT-153/+53 and of a proenkephalin gene reporter plasmid which contains a dimer of the enkephalin cAMP responsive element 2 (ENKCRE-2) in front of a minimal promoter was increased by cotransfection of a c-fos expression plasmid, indicating that nicotine may induce the proenkephalin gene in chromaffin cells via c-Fos which binds to the ENKCRE-2 element.
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Abbreviations
- API:
-
activator protein 1
- CAT:
-
chloramphenicol acetyl transferase
- CRE:
-
cAMP responsive element
- CREB:
-
cAMP response element binding protein
- DMEM:
-
Dulbecco's minimal essential medium
- ENKCRE:
-
enkaphalin cAMP responsive element
- IEG:
-
immediate early genes
- PENKCAT:
-
proenkephalin chloramphenicol acetyl transferase
References
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate phenol chloroform extraction. Anal Biochem 162:156–159
Farm CJ, Hollt V, Kley N (1990) Proenkephalin gene expression in cultured chromaffin cells is regulated at the transcriptional level. Prog Clin Biol Res 328:239–242
Farin CJ, Kley N, Höllt V (1990) Mechanisms involved in the transcriptional activation of proenkephalin gene expression in bovine chromaffin cells. J Biol Chem 265:19116–19121
Gandia L, Lopez MG, Fonteriz RI, Artalejo CR, Garcia AG (1987) Relative sensitivities of chromaffin cell calcium channels to organic and inorganic calcium antagonists. Neurosci Lett 77:333–338
Höllt V (1993) Regulation of opioid peptide gene expression. In: Herz A (ed) Handbook of experimental pharmacology: opioids I. Springer; Berlin Heidelberg New York, pp 307–346
Höllt V, Horn G (1991) Nicotine induces opioid peptide gene expression in hypothalamus and adrenal medulla of rats. In: Adlofer F; Thurau K (eds) Effects of nicotine on biological systems. Birkhäuser, Basel, pp 273–283
Höllt V, Horn G (1992) Effect of nicotine on mRNA levels encoding opioid peptides, vasopressin and alpha 3 nicotinic receptor subunit in the rat. Clin Investig 70:224–231
Huggenvik JI, Collard MW Stofko RE, Seasboltz AF, Uhler MD (1991) Regulation of the human enkephalin promoter by two isoforms of the catalytic subunit of cyclic adenosine 3′,5′-monophosphate-dependent protein kinase. Mol Endocrinol 5:921–930
Kley N (1988) Multiple regulation of proenkephalin gene expression by protein kinase. C J Biol Chem 263:2003–2008
Kley N, Loeffler JP, Pittius CW, Höllt V (1987) Involvement of ion channels in the induction of proenkephalin A gene expression by nicotine and CAMP in bovine chromaffin cells. J Biol Chem 262:4083–4089
Kobierski LA, Chu HM, Tan Y, Comb MJ (1991) cAMP-dependent regulation of proenkephalin by JunD and JunB: positive and negative effects of AP-1 proteins. Proc Natl Acad Sci USA 88:10222–10226
Konradi C, Kobierski LA, Nguyen TV, Heckers S, Hyman SE (1993) The cAMP-response-element-binding protein interacts, but Fos protein does not interact, with the proenkephalin enhancer in rat striatum Proc Natl Acad Sci USA 90:7005–7009
Masquilier D, Sassone Corsi P (1992) Transcriptional crosstalk: nuclear factors CREM and CREB bind to AP-1 sites and inhibit activation by Jun. J Biol Chem 267:22460–22466
Pittius CW, Kley N, Loeffler JP, Höllt V (1985) Quantitation of proenkephalin A messenger RNA in bovine brain, pituitary and adrenal medulla: correlation between mRNA and peptide levels. EMBO J 4:1257–1260
Rosen H, Douglass J, Herbert E (1984) Isolation and characterization of the rat proenkephalin gene. J Biol Chem 259:14309–14213
Sheng M, Dougan ST, McFadden G, Greenberg ME (1988) Calcium and growth factor pathways of c-fos transcriptional activation require distinct upstream regulatory sequences. Mol Cell Biol 8:2787–2796
Sheng M, Thompson MA, Greenberg ME (1991) CREB: a Ca(2+)-regulated transcription factor phosphorylated by calmodulin-dependent kinases. Science 252: 1427–1430
Sonnenberg JL, Rauscher FJ, Morgan JI, Curran T (1989) Regulation of proenkephalin by Fos and Jun. Science 246:1622–1625
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Wang, X., Bacher, B. & Höllt, V. Nicotine-induced gene expression of proenkephalin in bovine chromaffin cells. Clin Investig 72, 925–929 (1994). https://doi.org/10.1007/BF00190754
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DOI: https://doi.org/10.1007/BF00190754