GC factor 2 represses platelet-derived growth factor A-chain gene transcription and is itself induced by arterial injury.
Platelet-derived growth factor (PDGF) is a mitogen and chemoattractant for a wide variety of cell types. The genes encoding PDGF A chain (PDGF-A) and PDGF B chain (PDGF-B) reside on separate chromosomes and are independently regulated at the level of transcription. Regulatory events underlying inducible PDGF-A expression have been the focus of much investigation. However, mechanisms that inhibit transcription of this gene are not well understood. In this study, we report the capacity of a newly cloned DNA binding factor, GC factor 2 (GCF2), to repress expression driven by the human PDGF-A promoter. 5' Deletion and transient cotransfection analysis in vascular endothelial cells revealed that GCF2 repression is mediated by a nucleotide region located in the proximal region of the PDGF-A promoter. Electrophoretic mobility shift assays demonstrate that GCF2 binds to this region in a specific and dose-dependent manner. Interestingly, the site bound by GCF2 overlaps those for specificity protein-1 (Sp1) and early growth response factor-1 (Egr-1), zinc finger transcription factors that direct basal and inducible expression of the PDGF-A gene. Gel shift experiments revealed that GCF2 competes with these factors for interaction with the PDGF-A promoter. Overexpression of GCF2 suppressed endogenous PDGF-A expression in vascular endothelial cells and smooth muscle cells. GCF2 was induced on mechanical injury of cells in culture as well as after balloon injury of the rat carotid artery wall. Time course studies revealed the sustained induction of GCF2 after injury while PDGF-A levels sharply returned to baseline. Smooth muscle cell proliferation was inhibited by GCF2, an effect reversed by the addition of exogenous PDGF-AA. These findings demonstrate negative regulation of PDGF-A expression by GCF2. This is the first report of the induction of an endogenous transcriptional repressor in the rat vessel wall.
Khachigian
L M
LM
Centre for Thrombosis and Vascular Research, The University of New South Wales, Department of Haematology, Prince of Wales Hospital, Sydney, Australia. L.Khachigian@unsw.edu.au
Santiago
F S
FS
Rafty
L A
LA
Chan
O L
OL
Delbridge
G J
GJ
Bobik
A
A
Collins
T
T
Johnson
A C
AC
D000818:Animals; D001011:Aorta / Q000293:injuries / Q000473:pathology; D002417:Cattle; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics*; D004730:Endothelium, Vascular / Q000293:injuries* / Q000473:pathology; D051790:G-Box Binding Factors; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000293:injuries / Q000473:pathology; D010982:Platelet-Derived Growth Factor / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D012097:Repressor Proteins / Q000235:genetics*; D013314:Stress, Mechanical; D014157:Transcription Factors / Q000235:genetics*; D014158:Transcription, Genetic; D016335:Zinc Fingers
D004268:DNA-Binding Proteins; D051790:G-Box Binding Factors; D010982:Platelet-Derived Growth Factor; D012097:Repressor Proteins; D014157:Transcription Factors; C075278:platelet-derived growth factor A