title: Transforming growth factor β1 antagonizes the transcription, expression and vascular signaling of guanylyl cyclase/natriuretic peptide receptor A - role of δEF1.
abstract: The objective of this study was to determine the role of transforming growth factor β1 (TGF-β1) in transcriptional regulation and function of the guanylyl cyclase A/natriuretic peptide receptor A gene (Npr1) and whether cross-talk exists between these two hormonal systems in target cells. After treatment of primary cultured rat thoracic aortic vascular smooth muscle cells and mouse mesangial cells with TGF-β1, the Npr1 promoter construct containing a δ-crystallin enhancer binding factor 1 (δEF1) site showed 85% reduction in luciferase activity in a time- and dose-dependent manner. TGF-β1 also significantly attenuated luciferase activity of the Npr1 promoter by 62%, and decreased atrial natriuretic peptide-mediated relaxation of mouse denuded aortic rings ex vivo. Treatment of cells with TGF-β1 increased the protein levels of δEF1 by 2.4-2.8-fold, and also significantly enhanced the phosphorylation of Smad 2/3, but markedly reduced Npr1 mRNA and receptor protein levels. Over-expression of δEF1 showed a reduction in Npr1 promoter activity by 75%, while deletion or site-directed mutagenesis of δEF1 sites in the Npr1 promoter eliminated the TGF-β1-mediated repression of Npr1 transcription. TGF-β1 significantly increased the expression of α-smooth muscle actin and collagen type I α2 in rat thoracic aortic vascular smooth muscle cells, which was markedly attenuated by atrial natriuretic peptide in cells over-expressing natriuretic peptide receptor A. Together, the present results suggest that an antagonistic cascade exists between the TGF-β1/Smad/δEF1 pathways and Npr1 expression and receptor signaling that is relevant to renal and vascular remodeling, and may be critical in the regulation of blood pressure and cardiovascular homeostasis.
authors.affiliation: Department of Physiology, Tulane University Health Sciences Center and School of Medicine, New Orleans, LA, USA.
authors.forename: Anagha
authors.initials: A
authors.lastname: Sen
authors.affiliation: Department of Physiology, Tulane University Health Sciences Center and School of Medicine, New Orleans, LA, USA.
authors.forename: Prerna
authors.initials: P
authors.lastname: Kumar
authors.affiliation: Department of Physiology, Tulane University Health Sciences Center and School of Medicine, New Orleans, LA, USA.
authors.forename: Renu
authors.initials: R
authors.lastname: Garg
authors.affiliation: Department of Pharmacology, Tulane University Health Sciences Center and School of Medicine, New Orleans, LA, USA.
authors.forename: Sarah H
authors.initials: SH
authors.lastname: Lindsey
authors.affiliation: Department of Pharmacology, Tulane University Health Sciences Center and School of Medicine, New Orleans, LA, USA.
authors.forename: Prasad V G
authors.initials: PV
authors.lastname: Katakam
authors.affiliation: Department of Physiology, Tulane University Health Sciences Center and School of Medicine, New Orleans, LA, USA.
authors.forename: Meaghan
authors.initials: M
authors.lastname: Bloodworth
authors.affiliation: Department of Physiology, Tulane University Health Sciences Center and School of Medicine, New Orleans, LA, USA.
authors.forename: Kailash N
authors.initials: KN
authors.lastname: Pandey
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001011:Aorta / Q000166:cytology / Q000378:metabolism; D002460:Cell Line; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D005786:Gene Expression Regulation; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D008297:Male; D050527:Mesangial Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D009130:Muscle, Smooth / Q000166:cytology / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism; D051381:Rats; D017461:Receptors, Atrial Natriuretic Factor / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D051899:Smad2 Protein / Q000235:genetics* / Q000378:metabolism; D051900:Smad3 Protein / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D053773:Transforming Growth Factor beta1 / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D000071799:Zinc Finger E-box-Binding Homeobox 1
chemical_list: D000199:Actins; D024042:Collagen Type I; D018398:Homeodomain Proteins; D017461:Receptors, Atrial Natriuretic Factor; D051899:Smad2 Protein; D051900:Smad3 Protein; D014157:Transcription Factors; D053773:Transforming Growth Factor beta1; D000071799:Zinc Finger E-box-Binding Homeobox 1; C497784:Smad2 protein, rat; C497787:Smad3 protein, rat; C000630720:ZEB1 protein, rat; C498862:Acta2 protein, rat; C091715:atrial natriuretic factor receptor A
keywords: Smad; atrial natriuretic peptide; chromatin immunoprecipitation; gene expression; particulate guanylyl cyclase A