Stimulus-transcription Coupling in Pheochromocytoma Cells

To explore stimulus-transcription coupling in pheochromocytoma cells, we studied the biosynthetic response of chromogranin A, the major soluble protein co-stored and co-released with catecholamines, to chromaffin cells' physiologic nicotinic cholinergic secretory stimulation. Chromogranin A mRNA showed a time-dependent 3.87-fold response to nicotinic stimulation, and a nuclear run-off experiment indicated that the response occurred at a transcriptional level. Transfected chromogranin A promoter/luciferase reporter constructs were activated by nicotinic stimulation, in time- and dose-dependent fashions, in both rat PC12 pheochromocytoma cells and bovine chromaffin cells. Cholinergic subtype agents indicated that nicotinic stimulation was required. Promoter deletions established both positive and negative nicotinic response domains. Transfer of candidate promoter domains to a heterologous (thymidine kinase) promoter conferred region-specific nicotinic responses onto that promoter. A proximal promoter domain (from −93 to −62 base pairs) was activated in copy number- and distance-dependent fashion, and thus displayed features of a promoter element. Its activation was sufficient to account for the overall positive response to nicotine. Within this proximal region, a cAMP response element (CRE) was implicated as a major nicotinic response element, since a CRE point-gap mutation decreased nicotinic induction, transfer of CRE to a thymidine kinase promoter augmented the promoter's response to nicotine, and nicotine activated the CRE-binding protein CREB through phosphorylation at serine 133. We conclude that secretory stimulation of pheochromocytoma cells also activates the biosynthesis of the major secreted protein (chromogranin A), that the activation is transcriptional, and that a small proximal domain, including the CRE box, is, at least in part, both necessary and sufficient to account for the positive response to nicotine.

[1]  L. Eiden,et al.  Chromogranin A: current status as a precursor for bioactive peptides and a granulogenic/sorting factor in the regulated secretory pathway , 1995, Regulatory Peptides.

[2]  A. Laslop,et al.  Biosynthesis of large dense-core vesicles in PC12 cells: effects of depolarization and second messengers on the mRNA levels of their constituents. , 1995, Brain research. Molecular brain research.

[3]  D. O'Connor,et al.  Glucocorticoid activation of chromogranin A gene expression. Identification and characterization of a novel glucocorticoid response element. , 1994, The Journal of clinical investigation.

[4]  D. O'Connor,et al.  Cell type-specific gene expression in the neuroendocrine system. A neuroendocrine-specific regulatory element in the promoter of chromogranin A, a ubiquitous secretory granule core protein. , 1994, The Journal of clinical investigation.

[5]  A. S. Schneider,et al.  Nicotine Tolerance in Chromaffin Cell Cultures: Acute and Chronic Exposure to Smoking‐Related Nicotine Doses , 1994, Journal of neurochemistry.

[6]  G. Hendy,et al.  Human chromogranin A gene. Molecular cloning, structural analysis, and neuroendocrine cell-specific expression. , 1994, The Journal of biological chemistry.

[7]  B. Hiremagalur,et al.  Nicotine increases expression of tyrosine hydroxylase gene. Involvement of protein kinase A-mediated pathway. , 1993, The Journal of biological chemistry.

[8]  L. Helman,et al.  Secretory protein traffic. Chromogranin A contains a dominant targeting signal for the regulated pathway. , 1993, The Journal of clinical investigation.

[9]  T. Wagner,et al.  Gonadotropin secretion, synthesis, and gene expression in human growth hormone transgenic mice and in Ames dwarf mice. , 1993, Endocrinology.

[10]  M E Greenberg,et al.  Regulation of CREB phosphorylation in the suprachiasmatic nucleus by light and a circadian clock. , 1993, Science.

[11]  S. Hjorth,et al.  Serotonin 5‐HT1A Autoreceptor Blockade Potentiates the Ability of the 5‐HT Reuptake Inhibitor Citalopram to Increase Nerve Terminal Output of 5‐HT In Vivo: A Microdialysis Study , 1993, Journal of neurochemistry.

[12]  R. Kirchmair,et al.  Histamine induces a gene-specific synthesis regulation of secretogranin II but not of chromogranin A and B in chromaffin cells in a calcium-dependent manner. , 1993, The Journal of biological chemistry.

[13]  M. Brilliant,et al.  5' flanking sequences of the rat tyrosine hydroxylase gene target accurate tissue-specific, developmental, and transsynaptic expression in transgenic mice , 1992, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[14]  J. Mallet,et al.  AP‐1 complex and c‐fos transcription are involved in TPA provoked and trans‐synaptic inductions of the tyrosine hydroxylase gene: Insights into long‐term regulatory mechanisms , 1992, Journal of neuroscience research.

[15]  A. Mcmahon,et al.  Regulated expression of the tyrosine hydroxylase gene by membrane depolarization. Identification of the responsive element and possible second messengers. , 1992, The Journal of biological chemistry.

[16]  M. Grimes,et al.  The bovine chromogranin A gene: structural basis for hormone regulation and generation of biologically active peptides. , 1991, Molecular endocrinology.

[17]  D. O'Connor,et al.  Structure and function of the chromogranin A gene. Clues to evolution and tissue-specific expression. , 1991, The Journal of biological chemistry.

[18]  D. O'Connor,et al.  Suppression of chromogranin-A release from neuroendocrine sources in man: pharmacological studies. , 1991, The Journal of clinical endocrinology and metabolism.

[19]  K. W. Hasel,et al.  Nucleotide sequence of a cDNA coding for mouse cyclophilin , 1990, Nucleic Acids Res..

[20]  M. Stachowiak,et al.  Coordinate and differential regulation of phenylethanolamine N-methyltransferase, tyrosine hydroxylase and proenkephalin mRNAs by neural and hormonal mechanisms in cultured bovine adrenal medullary cells , 1990, Brain Research.

[21]  D. O'Connor,et al.  Chromogranin A. Storage and release in hypertension. , 1990, Hypertension.

[22]  M. Montminy,et al.  Characterization of a bipartite activator domain in transcription factor CREB , 1990, Cell.

[23]  A. Laslop,et al.  Insulin hypoglycemia increases the levels of neuropeptide Y and calcitonin gene-related peptide, but not of chromogranins A and B, in rat chromaffin granules , 1989, Regulatory Peptides.

[24]  D. O'Connor,et al.  Molecular Cloning of Chromogranin A From Rat Pheochromocytoma Cells , 1989, Hypertension.

[25]  D. Aunis,et al.  Effect of secretagogues on chromogranin A synthesis in bovine cultured chromaffin cells. Possible regulation by protein kinase C. , 1989, The Biochemical journal.

[26]  K. O’Malley,et al.  Characterization of rat and human tyrosine hydroxylase genes: functional expression of both promoters in neuronal and non-neuronal cell types. , 1988, Biochemical and biophysical research communications.

[27]  W. Roesler,et al.  Cyclic AMP and the induction of eukaryotic gene transcription. , 1988, The Journal of biological chemistry.

[28]  D. Aunis,et al.  Secretion from chromaffin cells is controlled by chromogranin A-derived peptides. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[29]  J. Northrop,et al.  Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[30]  D. Aunis,et al.  Chromogranin A Synthesis and Secretion in Chromaffin Cells , 1987, Journal of neurochemistry.

[31]  M. Karin,et al.  Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor , 1987, Cell.

[32]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[33]  H. Pollard,et al.  Contrasting monoamine oxidase activity and tyramine induced catecholamine release in PC12 and chromaffin cells , 1986, Neuroscience.

[34]  P. Tegtmeyer,et al.  Domain structure of the simian virus 40 core origin of replication , 1986, Molecular and cellular biology.

[35]  D. O'Connor,et al.  Immunological identification and characterization of chromogranins coded by poly(A) mRNA from bovine adrenal medulla and pituitary gland and human phaeochromocytoma. , 1984, The Journal of biological chemistry.

[36]  D. O'Connor,et al.  Chromogranin A, the major catecholamine storage vesicle soluble protein. Multiple size forms, subcellular storage, and regional distribution in chromaffin and nervous tissue elucidated by radioimmunoassay. , 1984, The Journal of biological chemistry.

[37]  A. Feinberg,et al.  A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. , 1983, Analytical biochemistry.

[38]  B. Howard,et al.  Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells , 1982, Molecular and cellular biology.

[39]  L. Greene,et al.  Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[40]  G. Bossi,et al.  Retinoic acid and cAMP differentially regulate human chromogranin A promoter activity during differentiation of neuroblastoma cells. , 1995, European journal of cancer.

[41]  B. Livett,et al.  Coordinate and differential regulation of proenkephalin A and PNMT mRNA expression in cultured bovine adrenal chromaffin cells: responses to secretory stimuli. , 1991, Brain Research. Molecular Brain Research.

[42]  D. O'Connor,et al.  Chromogranin A: posttranslational modifications in secretory granules. , 1991, Endocrinology.

[43]  D. O'Connor,et al.  Is physiologic sympathoadrenal catecholamine release exocytotic in humans? , 1990, Circulation.

[44]  P. Lelkes,et al.  Stimulus-secretion coupling in chromaffin cells , 1987 .