CAAT/enhancer-binding protein delta and cAMP-response element-binding protein mediate inducible expression of the nerve growth factor gene in the central nervous system.
Nerve growth factor (NGF) synthesis in the rat cerebral cortex is induced by the beta2-adrenergic receptor agonist clenbuterol (CLE). Because NGF is a crucial neurotrophic factor for basal forebrain cholinergic neurons, defining the mechanisms that regulate its transcription is important for developing therapeutic strategies to treat pathologies of these neurons. We previously showed that the transcription factor CCAAT/enhancer-binding protein delta (C/EBPdelta) contributes to NGF gene regulation. Here we have further defined the function of C/EBPdelta and identified a role for cAMP response element-binding protein (CREB) in NGF transcription. Inhibition of protein kinase A in C6-2B glioma cells suppressed CLE induction of an NGF promoter-reporter construct, whereas overexpression of protein kinase A increased NGF promoter activity, particularly in combination with C/EBPdelta. A CRE-like site that binds CREB was identified in the proximal NGF promoter, and C/EBPdelta and CREB were found to associate with the NGF promoter in vivo. Deletion of the CRE and/or C/EBP sites reduced CLE responsiveness of the promoter. In addition, ectopic expression of C/EBPdelta in combination with CLE treatment increased endogenous NGF mRNA levels in C6-2B cells. C/EBPdelta null mice showed complete loss of NGF induction in the cerebral cortex following CLE treatment, demonstrating a critical role for C/EBPdelta in regulating beta2-adrenergic receptor-mediated NGF expression in vivo. Thus, our findings demonstrate a critical role for C/EBPdelta in regional expression of NGF in the brain and implicate CREB in CLE-induced NGF gene transcription.
McCauslin
Christine Seitz
CS
Laboratory of Protein Dynamics and Signaling, National Cancer Institute, National Institutes of Health, Frederick, MD 21702-1201, USA.
Heath
Victoria
V
Colangelo
Anna Maria
AM
Malik
Radek
R
Lee
Sook
S
Mallei
Alessandra
A
Mocchetti
Italo
I
Johnson
Peter F
PF
D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000818:Animals; D001921:Brain / Q000502:physiology*; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000235:genetics / Q000378:metabolism; D051743:CCAAT-Enhancer-Binding Protein-delta / Q000235:genetics / Q000378:metabolism*; D050882:CREB-Binding Protein / Q000235:genetics / Q000378:metabolism*; D002976:Clenbuterol / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology; D005910:Glioma; D019426:Integrases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D016297:Mutagenesis, Site-Directed; D020932:Nerve Growth Factor / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D015533:Transcriptional Activation / Q000187:drug effects / Q000502:physiology*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
D000318:Adrenergic beta-Agonists; D022782:CCAAT-Enhancer-Binding Protein-beta; D012333:RNA, Messenger; D014764:Viral Proteins; D051743:CCAAT-Enhancer-Binding Protein-delta; D020932:Nerve Growth Factor; D050882:CREB-Binding Protein; C494364:Crebbp protein, mouse; D017868:Cyclic AMP-Dependent Protein Kinases; C045073:Cre recombinase; D019426:Integrases; D002976:Clenbuterol