Inducible platelet-derived growth factor D-chain expression by angiotensin II and hydrogen peroxide involves transcriptional regulation by Ets-1 and Sp1.
Platelet-derived growth factor D-chain (PDGF-D) is the newest member of the PDGF family of mitogens and chemoattractants expressed in a wide variety of cell types, including vascular smooth muscle cells (SMCs). The molecular mechanisms regulating PDGF-D transcription are not known. Primer extension analysis mapped a single transcriptional start site to the ccAGCGC motif with several potential Ets motifs located upstream. Ets-1, but not Ets-1 bearing only the DNA-binding domain, activates the PDGF-D promoter and mRNA expression in SMCs. Ets site D3 ((-470)GGAT(-467)) is singly required for basal and Ets-1-inducible PDGF-D promoter-dependent expression. D3 supports the interaction of endogenous and recombinant Ets-1 and Sp1. Sp1, like Ets-1, induces PDGF-D transcription and mRNA expression, which is blocked by mutant Ets-1. H2O2 stimulates Ets-1, but not Sp1, and activates D3-dependent PDGF-D transcription. Ets-1 and Sp1 siRNA block peroxide-inducible PDGF-D expression. Angiotensin II (ATII) induction of PDGF-D and Ets-1 was blocked by prior incubation of the cells with PEG-catalase, but not BSA, indicating that ATII-inducible Ets-1 and PDGF-D expression is mediated via H2O2. Thus, 2 separate trans-acting factors regulate PDGF-D transcription, alone and in response to oxidative stress.
Liu
Mary Yanxia
MY
Centre for Vascular Research, Department of Pathology, University of New South Wales, Sydney NSW 2052 Australia.
Eyries
Melanie
M
Zhang
Chunyan
C
Santiago
Fernando S
FS
Khachigian
Levon M
LM
D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D008222:Lymphokines / Q000235:genetics*; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D018384:Oxidative Stress; D010982:Platelet-Derived Growth Factor / Q000235:genetics*; D011401:Promoter Regions, Genetic; D050784:Proto-Oncogene Protein c-ets-1 / Q000235:genetics / Q000502:physiology*; D051381:Rats; D016329:Sp1 Transcription Factor / Q000502:physiology*; D024363:Transcription Initiation Site; D014158:Transcription, Genetic
C494043:Ets1 protein, rat; D008222:Lymphokines; C427493:PDGFD protein, human; D010982:Platelet-Derived Growth Factor; D050784:Proto-Oncogene Protein c-ets-1; D016329:Sp1 Transcription Factor; D000804:Angiotensin II; D006861:Hydrogen Peroxide