PMID- 8885243 OWN - NLM STAT- MEDLINE DCOM- 19970130 LR - 20211203 IS - 0888-8809 (Print) IS - 0888-8809 (Linking) VI - 10 IP - 9 DP - 1996 Sep TI - Stimulation of the P-450 side chain cleavage enzyme (CYP11A1) promoter through ras- and Ets-2-signaling pathways. PG - 1084-94 AB - Expression of the ovine P-450 side-chain cleavage enzyme gene (CYP11A1) is stimulated by epidermal growth factor (EGF) through a pathway that involves c-Jun in JEG-3 placental cells. Growth factor signaling involves ras-dependent and ras-independent signaling pathways, which in turn regulate gene transcription through related but distinct mitogen-activated protein kinase pathways (MAPKs) including the extracellular signal-regulated kinases (ERKs) and the stress-activated protein kinases (SAPKs). We investigated the intracellular signaling pathways governing EGF induction of the CYP11A1 promoter. EGF stimulation of the CYP11A1 promoter (4-fold) was reduced 60% by a dominant negative mutant of ras (N17), and 30-40% by antisense ras. EGF induced both ERK and SAPK activity in JEG-3 cells. EGF-induced CYP11A1 promoter activity was reduced 60% by the MEK1 inhibitor PD098059 and 50% by a dominant negative mutant of the ERK-specific regulator MEK1. In contrast, dominant negative mutants of the SAPK-specific activator, SEK1, induced a further increase in EGF-induced CYP11A1 promoter activity. Constitutively active mutants of ras (V12 or L61) increased CYP11A1 promoter activity 6- to 8-fold. Deletion of the EGF response element (EGF-RE) between -92 and -77 bp reduced ras induction by 60%; however, a residual 3-fold induction remained through the proximal -77 bp. Mutation of the EGF-RE AP-1-like sequence in the context of the native promoter reduced CYP11A1 promoter activation by ras 60%. The EGF-RE sequence was sufficient for 6-fold activation by ras in the context of an heterologous thymidine kinase promoter. Candidate transcription factor targets (c-Jun, c-Ets-2) for the ras-signaling cascade were examined for their effects on CYP11A1 promoter activity. Overexpression of c-Jun induced the CYP11A1 promoter through the EGF-RE; however, c-Ets-2 activation of the CYP11A1 promoter (12-fold) required the proximal ras-responsive promoter sequences that are distinct from the EGF/MEK/c-Jun-responsive element. Induction of the CYP11A1 promoter by EGF involves a ras/MEK1/AP-1-dependent pathway that is distinct from induction by ras/c-Ets-2. FAU - Pestell, R G AU - Pestell RG AD - Division of Endocrinology, Metabolism and Molecular Medicine, Northwestern University Medical School, Chicago, Illinois 60611, USA. FAU - Albanese, C AU - Albanese C FAU - Watanabe, G AU - Watanabe G FAU - Lee, R J AU - Lee RJ FAU - Lastowiecki, P AU - Lastowiecki P FAU - Zon, L AU - Zon L FAU - Ostrowski, M AU - Ostrowski M FAU - Jameson, J L AU - Jameson JL LA - eng GR - 5 T32 GM0852-10/GM/NIGMS NIH HHS/United States GR - HD-23519/HD/NICHD NIH HHS/United States GR - KO8 CA 620008 01/CA/NCI NIH HHS/United States PT - Journal Article PT - Research Support, Non-U.S. Gov't PT - Research Support, U.S. Gov't, P.H.S. PL - United States TA - Mol Endocrinol JT - Molecular endocrinology (Baltimore, Md.) JID - 8801431 RN - 0 (DNA-Binding Proteins) RN - 0 (ERF protein, human) RN - 0 (ETS2 protein, human) RN - 0 (Proto-Oncogene Protein c-ets-2) RN - 0 (Proto-Oncogene Proteins) RN - 0 (Repressor Proteins) RN - 0 (Trans-Activators) RN - 0 (Transcription Factor AP-1) RN - 0 (Transcription Factors) RN - 62229-50-9 (Epidermal Growth Factor) RN - EC 1.14.15.6 (Cholesterol Side-Chain Cleavage Enzyme) RN - EC 2.7.11.1 (Protein Serine-Threonine Kinases) RN - EC 2.7.11.17 (Calcium-Calmodulin-Dependent Protein Kinases) RN - EC 2.7.11.24 (JNK Mitogen-Activated Protein Kinases) RN - EC 2.7.11.24 (Mitogen-Activated Protein Kinases) RN - EC 2.7.11.25 (MAP Kinase Kinase Kinase 1) RN - EC 2.7.11.25 (MAP3K1 protein, human) SB - IM MH - Calcium-Calmodulin-Dependent Protein Kinases/genetics/metabolism MH - Cholesterol Side-Chain Cleavage Enzyme/drug effects/*genetics/*metabolism MH - Choriocarcinoma/genetics/metabolism/pathology MH - *DNA-Binding Proteins MH - Epidermal Growth Factor/metabolism/pharmacology MH - Gene Expression Regulation MH - Genes, jun MH - *Genes, ras MH - Humans MH - JNK Mitogen-Activated Protein Kinases MH - *MAP Kinase Kinase Kinase 1 MH - *Mitogen-Activated Protein Kinases MH - Promoter Regions, Genetic MH - Protein Serine-Threonine Kinases/genetics/metabolism MH - Proto-Oncogene Protein c-ets-2 MH - Proto-Oncogene Proteins/genetics/*metabolism MH - *Repressor Proteins MH - Sequence Deletion MH - Signal Transduction MH - Trans-Activators/genetics/metabolism MH - Transcription Factor AP-1/genetics/metabolism MH - *Transcription Factors MH - Transcription, Genetic/drug effects MH - Tumor Cells, Cultured EDAT- 1996/09/01 00:00 MHDA- 1996/09/01 00:01 CRDT- 1996/09/01 00:00 PHST- 1996/09/01 00:00 [pubmed] PHST- 1996/09/01 00:01 [medline] PHST- 1996/09/01 00:00 [entrez] AID - 10.1210/mend.10.9.8885243 [doi] PST - ppublish SO - Mol Endocrinol. 1996 Sep;10(9):1084-94. doi: 10.1210/mend.10.9.8885243.