A mechanistic role for DNA methylation in endothelial cell (EC)-enriched gene expression: relationship with DNA replication timing

Apurva V. Shirodkar, Rosanne St Bernard, Anna Gavryushova, Anna Kop, Britta J. Knight, Matthew Shu Ching Yan, Hon Sum Jeffrey Man, Maneesh Sud, Robert P. Hebbel, Peter Oettgen, William C. Aird, Philip A. Marsden

Research output: Contribution to journalArticlepeer-review

43 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)3531-3540
Number of pages10
JournalBlood
Volume121
Issue number17
DOIs
StatePublished - Apr 25 2013

Bibliographical note

Funding Information:
This work was supported by Canadian Institutes of Health grant MOP 79475 and National Heart, Lung, and Blood Institute grant PO1 HL076540-06A1 (P.A.M.). P.A.M. holds the Keenan Chair in Medical Research at St. Michael?s Hospital and the University of Toronto. A.V.S. is a recipient of the Canadian Institutes of Health Research Collaborative Graduate Training Program in Molecular Medicine award

Funding Information:
This work was supported by Canadian Institutes of Health grant MOP 79475 and National Heart, Lung, and Blood Institute grant PO1 HL076540-06A1 (P.A.M.). P.A.M. holds the Keenan Chair in Medical Research at St. Michael’s Hospital and the University of Toronto. A.V.S. is a recipient of the Canadian Institutes of Health Research Collaborative Graduate Training Program in Molecular Medicine award.

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