Ets-1 Stimulates Platelet-Derived Growth Factor A-Chain Gene Transcription and Vascular Smooth Muscle Cell Growth via Cooperative Interactions With Sp1
- 3 September 2004
- journal article
- Published by Ovid Technologies (Wolters Kluwer Health) in Circulation Research
- Vol. 95 (5), 479-487
- https://doi.org/10.1161/01.res.0000141135.36279.67
Abstract
The platelet-derived growth factor (PDGF) family of ligands (composed of A-, B-, C-, and D-chains), potent mitogens, and chemoattractants for cells of mesenchymal origin has been implicated in numerous vascular pathologies involving smooth muscle cell (SMC) hyperplasia. Understanding the molecular mechanisms mediating PDGF transcription would provide new insights into strategies to control PDGF-dependent pathophysiologic processes. We demonstrated previously that PDGF-A expression is under the positive regulatory influence of Sp1, Sp3, and Egr-1 and is negatively controlled by GCF2, NF-1(X), and WT-1. In this article, we demonstrate that Ets-1 induces PDGF-A expression in primary rat aortic SMCs at the level of transcription and mRNA expression. Electrophoretic mobility shift, supershift, and mutational analyses revealed a functional role for the −555TTCC−552 motif in the PDGF-A promoter that binds endogenous Ets-1. Chromatin immunoprecipitation analysis showed the interaction of endogenous and exogenous Ets-1 or glutathione S-transferase-tagged Ets-1, bearing only the DNA-binding domain with the authentic PDGF-A promoter. Conversely, dominant-negative mutant of Ets-1 blocked the promoter interaction of endogenous Ets-1. Overexpression of Ets-1 but not the mutant form of Ets-1 activates the PDGF-A promoter cooperatively with Sp1. Sp1, which interacts with Ets-1, failed to induce PDGF-A promoter-dependent expression if the promoter contained a site-specific mutation in this novel Ets-binding site. Small interfering RNA to Ets-1 and Sp1 blocked PDGF-BB- and serum-inducible PDGF-A expression. SMC growth was stimulated by Ets-1 and Sp1 separately and further increased by both factors together. Ets-1-inducible mitogenesis is blocked by antibodies neutralizing PDGF-A and involves activation of the PDGF α-receptor, which binds PDGF-A. These findings identify a functional cis-acting element for Ets-1 in the PDGF-A promoter and demonstrate that Sp1 and Ets-1 cooperatively activate PDGF-A transcription in vascular SMCs.Keywords
This publication has 26 references indexed in Scilit:
- Platelet-derived growth factor-BB and Ets-1 transcription factor negatively regulate transcription of multiple smooth muscle cell differentiation marker genesAmerican Journal of Physiology-Heart and Circulatory Physiology, 2004
- Regulation of Platelet-derived Growth Factor-A Chain by Krüppel-like Factor 5: NEW PATHWAY OF COOPERATIVE ACTIVATION WITH NUCLEAR FACTOR-κBPublished by Elsevier BV ,2004
- Ets-1 Protects Vascular Smooth Muscle Cells from Undergoing Apoptosis by Activating p21WAF1/Cip1Published by Elsevier BV ,2003
- Ets-1 Positively Regulates Fas Ligand Transcription via Cooperative Interactions with Sp1Published by Elsevier BV ,2002
- Peroxisome Proliferator–Activated Receptor-γ Ligands Inhibit Nuclear but Not Cytosolic Extracellular Signal–Regulated Kinase/Mitogen–Activated Protein Kinase–Regulated Steps in Vascular Smooth Muscle Cell MigrationJournal of Cardiovascular Pharmacology, 2001
- Induction of the Transcriptional Repressor Yin Yang-1 by Vascular Cell InjuryPublished by Elsevier BV ,2001
- Angiotensin II (ATII)-inducible Platelet-derived Growth Factor A-chain Gene Expression Is p42/44 Extracellular Signal-regulated Kinase-1/2 and Egr-1-dependent and Mediated via the ATII Type 1 but Not Type 2 ReceptorJournal of Biological Chemistry, 1999
- Induction of Ets-1 in endothelial cells during reendothelialization after denuding injuryJournal of Cellular Physiology, 1998
- Interplay of Sp1 and Egr-1 in the Proximal Platelet-derived Growth Factor A-Chain Promoter in Cultured Vascular Endothelial CellsJournal of Biological Chemistry, 1995
- Production of Platelet-Derived Growth Factor–like Mitogen by Smooth-Muscle Cells from Human AtheromaNew England Journal of Medicine, 1988