The cAMP-response element mediates induction of secretogranin II by CHX and FSK in GH4C1 cells.
The effect of cAMP on secretogranin II (SgII) gene transcription in GH4C1 (GH) cells is not observed unless protein synthesis is inhibited. We have defined elements in the SgII promoter that mediate regulation by cycloheximide (CHX) and forskolin (FSK) and characterized the nuclear proteins that interact with them. GH cells were transfected with p2774Luc, p351Luc, p242Luc, and p223Luc containing 2,612, 189, 80, and 61 bp of the SgII promoter upstream of the luciferase gene, respectively. Treatment with CHX and FSK increased promoter activity 8- to 12-fold in cells transfected with p2774Luc, p351Luc, and p242Luc but had not effect in cells transfected with p223Luc. The same 19-bp element (-80 to -62) mediates regulation by CHX alone, as CHX caused a 3.8-fold increase in activity in GH cells transfected with p242Luc but not p223Luc. Gel mobility shifts using sequences -84 to -53 resulted in three complexes, which contained cAMP response element-binding protein heterodimerized with cAMP response element modulator or activating transcription factor-1. No differences were observed in complex formation when cells were treated with either CHX, FSK, or CHX and FSK. Thus CHX affects the response to FSK in GH cells by inhibiting the synthesis of a protein, which does not itself interact with DNA or affect the binding of CRE-binding proteins with the SgII promoter, but likely interferes with the interaction of CRE-binding proteins with the general transcriptional machinery.
Jones
L C
LC
Department of Pharmacology, College of Medicine, University of South Alabama, Mobile 36688, USA.
Scammell
J G
JG
D000818:Animals; D002864:Chromogranins; D005576:Colforsin / Q000494:pharmacology*; D017362:Cyclic AMP Response Element-Binding Protein / Q000502:physiology*; D003513:Cycloheximide / Q000494:pharmacology*; D017353:Gene Deletion; D015870:Gene Expression / Q000187:drug effects; D005786:Gene Expression Regulation / Q000502:physiology; D009490:Neurosecretory Systems / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000187:drug effects; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D014407:Tumor Cells, Cultured
D002864:Chromogranins; D017362:Cyclic AMP Response Element-Binding Protein; D009687:Nuclear Proteins; D011500:Protein Synthesis Inhibitors; D011506:Proteins; D005576:Colforsin; D003513:Cycloheximide