The Korean Journal of Physiology and Pharmacology

The Korean Society of Pharmacology (대한약리학회)
권호 표지

PKA-Mediated Regulation of B/K Gene Transcription in PC12 Cells

  • Choi, Mi-Hyun (Department of Biochemistry, The Catholic University of Korea College of Medicine) ;
  • Kim, Ho-Shik (Department of Biochemistry, The Catholic University of Korea College of Medicine) ;
  • Choi, Sung-Ho (R&D Center, PetaGen, B199F Yonsei Engineering Research Complex) ;
  • Kim, Mi-Young (Dae-Myung Science) ;
  • Jang, Yoon-Seong (Department of Biochemistry, The Catholic University of Korea College of Medicine) ;
  • Jang, Young-Min (Department of Biochemistry, The Catholic University of Korea College of Medicine) ;
  • Lee, Jeong-Hwa (Department of Biochemistry, The Catholic University of Korea College of Medicine) ;
  • Jeong, Seong-Whan (Department of Biochemistry, The Catholic University of Korea College of Medicine) ;
  • Kim, In-Kyung (Department of Biochemistry, The Catholic University of Korea College of Medicine) ;
  • Kwon, Oh-Joo (Department of Biochemistry, The Catholic University of Korea College of Medicine)
  • Published : 2005.12.21
Download PDF
⟨ Previous Next ⟩

Abstract

B/K protein is a novel protein containing double C2-like domains. We examined the specific signaling pathway that regulates the transcription of B/K in PC12 cells. When the cells were treated with forskolin ($50{\mu}M$), B/K mRNA and protein levels were time-dependently decreased, reaching the lowest level at 3 or 4 hr, and thereafter returning to the control level. Chemicals such as dibutyryl-cAMP, cellpermeable cyclic AMP (cAMP) analogue and CGS21680, adenosine receptor $A_{2A}$ agonist, also repressed the B/K transcription. However, 1,9-dideoxyforskolin did not show inhibitory effect on B/K transcription, suggesting direct involvement of cAMP in the forskolin-induced inhibition of B/K transcription. Effect of forskolin, dibutyryl cAMP and CGS21680 was significantly reduced in PKA-deficient PC12 cell line (PC12-123.7). One cAMP-response element (CRE)-like sequence (B/K CLS) was found in the promoter region of B/K DNA, and electrophoretic mobility shift assay indicated its binding to CREM and CREB. Forskolin significantly suppressed the promoter activity in CHO-K1 cells transfected with the constructs containing B/K CLS, but not with the construct in which B/K CLS was mutated (AC：TG). Taken together, we suggest that the transcription of B/K gene in PC12 cells may be regulated by PKA-dependent mechanism.

Keywords

References

  1. Agnihotri N, Kisaalita WS, Keith CH. Free cyclic AMP increases in PC12 cells on depolarization. J Neurosci Res 47: 555-560, 1997 https://doi.org/10.1002/(SICI)1097-4547(19970301)47:5<555::AID-JNR11>3.0.CO;2-X
  2. Brose N, Petrenko AG, Sdhof TC, Jahn R. Synaptotagmin: a calcium sensor on the synaptic vesicle surface. Science 256: 1021- 1025, 1992 https://doi.org/10.1126/science.1589771
  3. Ding WQ, Dong M, Ninova D, Holicky EL, Stegall MD, Miller LJ. Forskolin suppresses insulin gene transcription in islet -cells through a protein kinase A-independent pathway. Cell Signal 15: 27-35, 2002 https://doi.org/10.1016/S0898-6568(02)00051-7
  4. Fass DM, Butler EF, Goodman H. Deacetylase activity is required for cAMP activation of subset of CREB target genes. J Biol Chem 278: 43014-43019, 2003 https://doi.org/10.1074/jbc.M305905200
  5. Fitzgerald LR, Li Z, Machida CA, Fishman PH, Duman RS. Adrenergic regulation of ICER (inducible cyclic AMP early repressor) and beta1-adrenergic receptor gene expression in C6 glioma cells. J Neurochem 67: 490-497, 1996 https://doi.org/10.1046/j.1471-4159.1996.67020490.x
  6. Foulkes NS, Duval G, Sassone-Corsi P. Adaptive inducibility of CREM as transcriptional memory of circadian rhythms. Nature 381: 83-85, 1996 https://doi.org/10.1038/381083a0
  7. Ginty DD, Glowacka D, DeFranco C, Wagner JA. Nerve growth factor-induced neuronal differentiation after dominant repression of both type I and type II cAMP-dependent protein kinase activities. J Biol Chem 266: 15325-15333, 1991
  8. Hansen TV, Rehfeld JF, Nielsen FC. KCl and forskolin synergistically up-regulate cholecystokinin gene expression via coordinate activation of CREB and the co-activator CBP. J Neurochem 89: 15-23, 2004 https://doi.org/10.1046/j.1471-4159.2003.02252.x
  9. Hoshi T, Garber SS, Aldrich RW. Effect of forskolin on voltagegated $K^+$ channels is independent of adenylate cyclase activation. Science 240: 1652-1655, 1988 https://doi.org/10.1126/science.2454506
  10. Ibata K, Fukuda M, Mikoshiba K. Inositol 1, 3, 4, 5-tetrakisphosphate binding activities of neuronal and non-neuronal synaptotagmins. J Biol Chem 273: 12267-12273, 1998 https://doi.org/10.1074/jbc.273.20.12267
  11. Jang YS, Lee MY, Choi SH, Kim MY, Chin H, Jeong SW, Kim IK, Kwon OJ. Expression of B/K protein in the hippocampus of kainate-induced rat seizure model. Brain Res 999: 203-211, 2004 https://doi.org/10.1016/j.brainres.2003.11.047
  12. Ju WK, Choi SH, Kwon JS, Kwon OJ, Lee MY, Oh SJ, Moon JL, Chun MH. Expression of brain/kidney protein in Muller cells of rat retina following transient ischemia. Neurosci Lett 293: 53- 56, 2000 https://doi.org/10.1016/S0304-3940(00)01492-0
  13. Krueger DA, Mao D, Warner EA, Dowd DR. Functional analysis of the mouse ICER (Inducible cAMP Early Repressor) promoter: evidence for a protein that blocks calcium responsiveness of the CAREs (cAMP autoregulatory elements). Mol Endocrinol 13: 1207-1217, 1999 https://doi.org/10.1210/me.13.7.1207
  14. Kwon OJ, Gainer H, Wray S, Chin H. Identification of a novel protein containing two C2 domains selectively expressed in the rat brain and kidney. FEBS Lett 378: 135-139, 1996 https://doi.org/10.1016/0014-5793(95)01430-6
  15. Kwon OJ, Ju WK, Choi SH, Lee MY, Oh SJ, Chun MH. The expression and cellular localization of brain/kidney (B/K) protein in the rat retina. Brain Res 860: 178-180, 2000 https://doi.org/10.1016/S0006-8993(00)02014-X
  16. Lao G, Scheuss V, Gerwin CM, Su Q, Mochida S, Rettig J, Sheng ZH. Syntaphilin: a syntaxin-1 clamp that controls SNARE assembly. Neuron 25: 191-201, 2000 https://doi.org/10.1016/S0896-6273(00)80882-X
  17. Lee MY, Choi SH, Shin SL, Chin H, Kwon OJ. Distribution of B/K protein in rat brain. Cell Tissue Res 303: 47-56, 2001 https://doi.org/10.1007/s004410000221
  18. Molina CA, Foulkes NS, Lalli E, Sassone-Corsi P. Inducibility and negative autoregulation of CREM: an alternative promoter directs the expression of ICER, an early response repressor. Cell 75: 875-886, 1993 https://doi.org/10.1016/0092-8674(93)90532-U
  19. Monaco L, Sassone-Corsi P. Cross-talk in signal transduction: Rasdependent induction of cAMP-responsive transcriptional repressor ICER by nerve growth factor. Oncogene 15: 2493-2500, 1997 https://doi.org/10.1038/sj.onc.1201636
  20. Montminy M. Transcriptional regulation by cyclic AMP. Annu Rev Biochem 66: 807-822, 1997 https://doi.org/10.1146/annurev.biochem.66.1.807
  21. Mukhopadhyay P, Roy KB. Protein engineering of BamHI restriction endonuclease: replacement of Cys54 by Ala enhances catalytic activity. Prot Engineer 11: 931-935, 1998 https://doi.org/10.1093/protein/11.10.931
  22. Nakamura H, Watanabe T, Kakiuchi S, Wada H. Enzymatic cleavage of the O2'-butyryl ester bond of N6, O2'-dibutyryl cyclic AMP and its possible involvement in the action of cyclic AMP derivatives in vivo. J Biochem 85: 1321-1329, 1979
  23. Poppe M, Hahm B, Eickelbaum W, Arand M, Paweletz N, Knehr M. Use of PCR to screen for promoter elements in genomic DNA library clones. Biotechniques 26: 718-722, 1999
  24. Saini HS, Gorse KM, Boxer LM, Sato-Bigbee C. Neurotrophin-3 and a CREB-mediated signaling pathway regulate Bcl-2 expression in oligodendrocyte progenitor cells. J Neurochem 89: 951-961, 2004 https://doi.org/10.1111/j.1471-4159.2004.02365.x
  25. Saitoh O, Saitoh Y, Nakata H. Regulation of A2A adenosine receptor mRNA expression by agonists and forskolin in PC12 cells. Neuroreport 5: 1317-1320, 1994 https://doi.org/10.1097/00001756-199406270-00005
  26. Scheibe RJ, Ginty DD, Wagner JA. Retinoic acid stimulates the differentiation of PC12 cells that are deficient in cAMPdependent protein kinase. J Cell Biol 113: 1173-1182, 1991 https://doi.org/10.1083/jcb.113.5.1173
  27. Servillo G, Penna L, Foulkes NS, Magni MV, Della Fazia MA, Sassone-Corsi P. Cyclic AMP signalling pathway and cellular proliferation: induction of CREM during liver regeneration. Oncogene 14: 1601-1606, 1997 https://doi.org/10.1038/sj.onc.1200996
  28. Shao X, Davletov BA, Sutton RB, Sudhof TC, Rizo J. Bipartite $Ca^{2+}$- binding motif in C2 domains of synaptotagmin and protein kinase C. Science 273: 248-251, 1996 https://doi.org/10.1126/science.273.5272.248
  29. Sheng M, McFadden G, Greenberg ME. Membrane depolarization and calcium induce c-fos transcription via phosphorylation of transcription factor CREB. Neuron 4: 571-582, 1990 https://doi.org/10.1016/0896-6273(90)90115-V
  30. Sheng M, Thompson MA, Greenberg ME. CREB: a $Ca^{2+}$-regulated transcription factor phosphorylated by calmodulin-dependent kinases. Science 252: 1427-1430, 1991 https://doi.org/10.1126/science.1646483
  31. Stehle JH, Foulkes NS, Molina CA, Simonneaux V, Pevet P, Sassone-Corsi P. Adrenergic signals direct rhythmic expression of transcriptional repressor CREM in the pineal gland. Nature 365: 314-320, 1993. https://doi.org/10.1038/365314a0
  32. Sudhof TC, Rizo J. Synaptotagmins: C2-domain proteins that regulate membrane traffic. Neuron 17: 379-388, 1996 https://doi.org/10.1016/S0896-6273(00)80171-3
  33. Thommesen L, Norsett K, Sandvik AK, Hofsli E, Laegreid A. Regulation of inducible cAMP early repressor expression by gastrin and cholecystokinin in the pancreatic cell line AR42J. J Biol Chem 275: 4244-4250, 2000 https://doi.org/10.1074/jbc.275.6.4244
  34. Tigyi G, Fischer DJ, Sebok A, Marshall F, Dyer DL, Miledi R. Lysophosphatidic acid-induced neurite retraction in PC12 cells: neurite-protective effects of cyclic AMP signaling. J Neurochem 66: 549-558, 1996 https://doi.org/10.1046/j.1471-4159.1996.66020549.x
  35. Tinti C, Conti B, Cubells JF, Kim KS, Baker H, Joh TH. Inducible cAMP early repressor can modulate tyrosine hydroxylase gene expression after stimulation of cAMP synthesis. J Biol Chem 271: 25375-25381, 1996 https://doi.org/10.1074/jbc.271.41.25375
  36. Tocco G, Bi X, Vician L, Lim IK, Herschman H, Baudry M. Two synaptotagmin genes, Syt1 and Syt4, are differentially regulated in adult brain and during postnatal development following kainic acid-induced seizures. Brain Res Mol Brain Res 40: 229- 239, 1996 https://doi.org/10.1016/0169-328X(96)00055-1
  37. Van Nguyen T, Kobierski L, Comb M, Hyman SE. The effect of depolarization on expression of the human proenkephalin gene is synergistic with cAMP and dependent upon a cAMP-inducible enhancer. J Neurosci 10: 2825-2833, 1990
  38. Vician L, Lim IK, Ferguson G, Tocco G, Baudry M, Herschman HR. Synaptotagmin IV is an immediate early gene induced by depolarization in PC12 cells and in brain. Proc Natl Acad Sci USA 92: 2164-2168, 1995 https://doi.org/10.1073/pnas.92.6.2164
  39. Wagoner PK, Pallotta BS. Modulation of acetylcholine receptor desensitization by forskolin is independent of cAMP. Science 240: 1655-1657, 1988 https://doi.org/10.1126/science.2454507
  40. Zhang X, Odom DT, Koo SH, Conkright MD, Canettieri G, Best J, Chen H, Jenner R, Herbolsheimer E, Jacobsen E, Kadam S, Ecker JR, Emerson B, Hogenesch JB, Unterman T, Young RA,Montminy M. Genome-wide analysis of cAMP-response element binding protein occupancy, phosphorylation, and target gene activation in human tissues. Proc Natl Acad Sci USA 102: 4459-4464, 2005 https://doi.org/10.1073/pnas.0501076102