Year: 2019
Type: dissertation
Abstract: Endothelial cells (ECs) have distinct gene expression and morphology depending on their location in the vasculature. A prevailing assumption is that these differences are largely caused by environmental factors, which differentially modulate gene expression in an otherwise homogeneous cell population. Here, we investigated the mechanistic basis for heterogeneity in EC gene expression and its functional importance, using P-selectin as a model gene. We found that heterogeneity exists basally in P-selectin protein expression across several human EC types. RNA FISH also showed heterogeneity in SELP transcription in isogenic populations of human umbilical vein endothelial cells (HUVEC). Differential proximal promoter DNA methylation, in part, accounted for this protein and RNA heterogeneity. In vitro methylation of the SELP promoter repressed transcription in episomal promoter reporter transfections. Importantly, promoter methylation was shown to be mitotically heritable. Using informative heterozygous SNPs in the SELP gene, we also report that P-selectin is monoallelically expressed in HUVEC.
Source: TSpace (University of Toronto)
Author: Lucy Chen
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2964752832