title: A mechanistic role for DNA methylation in endothelial cell (EC)-enriched gene expression: relationship with DNA replication timing.
abstract: Proximal promoter DNA methylation has been shown to be important for regulating gene expression. However, its relative contribution to the cell-specific expression of endothelial cell (EC)-enriched genes has not been defined. We used methyl-DNA immunoprecipitation and bisulfite conversion to analyze the DNA methylation profile of EC-enriched genes in ECs vs nonexpressing cell types, both in vitro and in vivo. We show that prototypic EC-enriched genes exhibit functional differential patterns of DNA methylation in proximal promoter regions of most (eg, CD31, von Willebrand factor [vWF], VE-cadherin, and intercellular adhesion molecule-2), but not all (eg, VEGFR-1 and VEGFR-2), EC-enriched genes. Comparable findings were evident in cultured ECs, human blood origin ECs, and murine aortic ECs. Promoter-reporter episomal transfection assays for endothelial nitric oxide synthase, VE-cadherin, and vWF indicated functional promoter activity in cell types where the native gene was not active. Inhibition of DNA methyltransferase activity indicated important functional relevance. Importantly, profiling DNA replication timing patterns indicated that EC-enriched gene promoters with differentially methylated regions replicate early in S-phase in both expressing and nonexpressing cell types. Collectively, these studies highlight the functional importance of promoter DNA methylation in controlling vascular EC gene expression.
authors: Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada.
authors: Apurva V
authors: AV
authors: Shirodkar
authors: Rosanne
authors: R
authors: St Bernard
authors: Anna
authors: A
authors: Gavryushova
authors: Anna
authors: A
authors: Kop
authors: Britta J
authors: BJ
authors: Knight
authors: Matthew Shu-Ching
authors: MS
authors: Yan
authors: Hon-Sum Jeffrey
authors: HS
authors: Man
authors: Maneesh
authors: M
authors: Sud
authors: Robert P
authors: RP
authors: Hebbel
authors: Peter
authors: P
authors: Oettgen
authors: William C
authors: WC
authors: Aird
authors: Philip A
authors: PA
authors: Marsden
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics; D001011:Aorta / Q000166:cytology / Q000378:metabolism; D015820:Cadherins / Q000235:genetics; D002417:Cattle; D015815:Cell Adhesion Molecules / Q000235:genetics; D002478:Cells, Cultured; D047369:Chromatin Immunoprecipitation; D019175:DNA Methylation; D042522:DNA Replication Timing; D020405:Dermis / Q000166:cytology / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism; D005786:Gene Expression Regulation; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D051379:Mice; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D016196:S Phase / Q000502:physiology*; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics; D014841:von Willebrand Factor / Q000235:genetics; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D015815:Cell Adhesion Molecules; D052250:Nitric Oxide Synthase Type III; D040281:Vascular Endothelial Growth Factor Receptor-1; D040301:Vascular Endothelial Growth Factor Receptor-2; D014841:von Willebrand Factor; D000099152:Cadherin 5