title: Neurotrophin activation of catecholamine storage vesicle protein gene expression: signaling to chromogranin a biosynthesis.
abstract: Nerve growth factor differentiates precursor cells into sympathetic neurons. Does acquisition of a "neuronal" phenotype after nerve growth factor involve biosynthesis of chromogranin A, the major soluble protein in chromaffin granule cores? Nerve growth factor activated chromogranin A gene expression 7.6-fold in PC12 pheochromocytoma cells, and similarly activated PC12-transfected mouse, rat or human chromogranin A promoter/reporter constructs. Chromogranin A promoter 5'-deletions narrowed the nerve growth factor response element to a region from - 77 to - 61 bp upstream of the cap site, a region containing the chromogranin A cyclic AMP response element (TGACGTAA). Three different site-directed mutations of the cyclic AMP response element each reduced the nerve growth factor effect by >90%. Transfer of the cyclic AMP response element to a heterologous (thymidine kinase) promoter activated that promoter approximately 5-fold after nerve growth factor, while transfer of a cyclic AMP response element point-gap mutant (TGA-GTAA) to a heterologous promoter abolished the nerve growth factor effect. These findings indicate that the cyclic AMP response element in cis is, at least in part, both necessary and sufficient to activate the chromogranin A gene. Chemical blockade of the nerve growth factor receptor TrkA or the mitogen-activated protein kinase pathway component MEK substantially diminished nerve growth factor-induced expression of chromogranin A. By contrast, the response of chromogranin A to nerve growth factor was not impaired after blockade of phospholipase C-gamma or phosphoinositide-3 kinase. Chemical blockade of TrkA, Ras, MEK or mitogen-activated protein kinase similarly inhibited nerve growth factor activation of chromogranin A. Expression of constitutively activated Ras, Raf or MEK mutants increased chromogranin A promoter activity. Expression of dominant negative (inhibitory) mutants of Sos, Ha-Ras, Rafl, mitogen-activated protein kinase, ribosomal protein S6 serine kinase II (CREB kinase) or CREB (KCREB) each inhibited the nerve growth factor-induced increase in chromogranin A promoter activity. Thus, each component of the mitogen-activated protein kinase pathway is crucially involved in relaying the nerve growth factor signal in trans to the chromogranin A gene, in the following proposed sequence: nerve growth factor --> TrkA --> Shc/Grb2/Sos --> Ras --> Raf --> MEK --> mitogen-activated protein kinase --> ribosomal protein S6 serine kinase II --> CREB cyclic AMP response element.
authors: Department of Medicine and Center for Molecular Genetics, University of California, San Diego, USA.
authors: S K
authors: SK
authors: Mahata
authors: M
authors: M
authors: Mahata
authors: H
authors: H
authors: Wu
authors: R J
authors: RJ
authors: Parmer
authors: D T
authors: DT
authors: O'Connor
mesh_terms: D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D002227:Carbazoles; D002395:Catecholamines / Q000378:metabolism*; D020843:Chamomile; D019439:Chromaffin Cells / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology; D002837:Chromaffin Granules / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D053379:Chromogranin A; D002864:Chromogranins / Q000096:biosynthesis / Q000235:genetics*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D017353:Gene Deletion; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology; D007211:Indoles; D008148:Lovastatin / Q000494:pharmacology; D051379:Mice; D020929:Mitogen-Activated Protein Kinase Kinases; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000502:physiology; D009414:Nerve Growth Factors / Q000494:pharmacology*; D009822:Oils, Volatile / Q000494:pharmacology; D016716:PC12 Cells; D019696:Peptidylprolyl Isomerase / Q000235:genetics; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010946:Plants, Medicinal; D011401:Promoter Regions, Genetic / Q000502:physiology; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics; D020917:Receptor, trkA; D017475:Receptors, Nerve Growth Factor / Q000235:genetics; D014158:Transcription, Genetic / Q000502:physiology; D014162:Transfection; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D000470:Alkaloids; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002227:Carbazoles; D002395:Catecholamines; D053379:Chromogranin A; D002864:Chromogranins; D004791:Enzyme Inhibitors; D005419:Flavonoids; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D007211:Indoles; D008148:Lovastatin; D020929:Mitogen-Activated Protein Kinase Kinases; D009414:Nerve Growth Factors; D009822:Oils, Volatile; D019696:Peptidylprolyl Isomerase; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D020794:Receptor Protein-Tyrosine Kinases; D020917:Receptor, trkA; D017475:Receptors, Nerve Growth Factor; D010738:Type C Phospholipases; C506150:CHGA protein, human; C506155:chromogranin A, mouse; C506153:chromogranin A, rat; C063769:KT 5926; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one