PC12 cells regulate inducible cyclic AMP (cAMP) element repressor expression to differentially control cAMP response element-dependent transcription in response to nerve growth factor and cAMP.
Both cyclic AMP (cAMP) and nerve growth factor (NGF) have been shown to cause rapid activation of cAMP response element-binding protein (CREB) by phosphorylation of serine 133, but additional regulatory events contribute to CREB-targeted gene expression. Here, we have used stable transfection with a simple cAMP response element (CRE)-driven reporter to address the kinetics of CRE-dependent transcription during neuronal differentiation of PC12 cells. In naive cells, dibutyryl cAMP (dbcAMP) generated a rapid increase in CRE-driven luciferase activity by 5 h that returned to naive levels by 24 h. Luciferase induction after NGF treatment was delayed until 48 h when CRE-driven luciferase expression became TrkA dependent. Blocking histone deacetylase (HDAC) activity accelerated NGF-dependent CRE-driven luciferase expression by at least 24 h and resulted in a sustained cAMP-dependent expression of CRE-driven luciferase beyond 24 h. Inhibition of protein synthesis before stimulation with NGF or dbcAMP indicated that both stimuli induce expression of a transcriptional repressor that delays NGF-dependent and attenuates cAMP-dependent CRE-driven transcription. NGF caused a rapid but transient HDAC-dependent increase in inducible cAMP element repressor (ICER) expression, but ICER expression was sustained with increased cAMP. Depletion of ICER from PC12 cells indicated that HDAC-dependent ICER induction is responsible for the delay in CRE-dependent transcription after NGF treatment.
Chang
Jay H
JH
Cellular and Molecular Pathology Graduate Program, David Geffen School of Medicine, University of California, Los Angeles, California, USA.
Vuppalanchi
Deepika
D
van Niekerk
Erna
E
Trepel
Jane B
JB
Schanen
N Carolyn
NC
Twiss
Jeffery L
JL
D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002227:Carbazoles / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D047369:Chromatin Immunoprecipitation / Q000379:methods; D000242:Cyclic AMP / Q000494:pharmacology*; D051786:Cyclic AMP Response Element Modulator / Q000494:pharmacology*; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism*; D004347:Drug Interactions; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects* / Q000502:physiology; D005786:Gene Expression Regulation / Q000187:drug effects*; D047468:Immunoprecipitation / Q000379:methods; D026121:Indole Alkaloids; D008156:Luciferases / Q000378:metabolism; D020932:Nerve Growth Factor / Q000494:pharmacology*; D016716:PC12 Cells / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013997:Time Factors; D014162:Transfection / Q000379:methods
D002227:Carbazoles; D017362:Cyclic AMP Response Element-Binding Protein; D004791:Enzyme Inhibitors; D026121:Indole Alkaloids; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051786:Cyclic AMP Response Element Modulator; D003994:Bucladesine; D020932:Nerve Growth Factor; C049985:staurosporine aglycone; D000242:Cyclic AMP; D008156:Luciferases