DEPLETION OF DNMT1 IN DIFFERENTIATED HUMAN CELLS HIGHLIGHTS KEY CLASSES OF DEPENDENT GENES

Karla O'Neill, Rachelle E Irwin, Sarah-Jayne Mackin, Avinash Thakur, Sara-Jayne Thursby, Ciske Bertens, Laura Masala, Jayne E.P. Loughery, Darragh G McArt, CP Walsh

Research output: Contribution to conferenceAbstract

Abstract

DNA methylation is a critical mechanism for regulating gene expression and ensuring genomic stability. However, loss of function mutations of methyltransferase enzymes such as DNMT1 in normal differentiated cells result in a lethal phenotype. Consequently, existing investigations have only assessed DNMT1 knockdowns in embryonic stem cells or cancer cell lines. Here, isogenic lines of hypomorphic, normal, immortalised fibroblasts have instead been generated via stable integration with short hairpin RNA. Enrichment analysis of epigenome-wide methylation arrays indicated widespread demethylation within promoter and gene body regions. In addition, four specific gene categories were highlighted as most affected; protocadherins, genes regulating body mass, olfactory receptors and cancer/testis antigens. Comparison of short-term siRNA and long-term shRNA-mediated depletion of DNMT1 indicated that many regions recover methylation as shRNA-containing cell lines adapt to lowered levels of DNMT1. Interestingly, polycomb-regulated genes are refractory to de novo DNA methylation in these cells following recovery, reinforcing the concept of mutually-exclusive domains that are regulated by these two major epigenetic mechanisms.
Original languageEnglish
Pages54-64
Publication statusPublished - 2018
Event20th Meeting of the Irish Society of Human Genetics - Croke Park, Dublin, Dublin, Ireland
Duration: 15 Sep 201715 Sep 2018
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5849961/

Conference

Conference20th Meeting of the Irish Society of Human Genetics
CountryIreland
CityDublin
Period15/09/1715/09/18
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O'Neill, K., Irwin, R. E., Mackin, S-J., Thakur, A., Thursby, S-J., Bertens, C., ... Walsh, CP. (2018). DEPLETION OF DNMT1 IN DIFFERENTIATED HUMAN CELLS HIGHLIGHTS KEY CLASSES OF DEPENDENT GENES. 54-64. Abstract from 20th Meeting of the Irish Society of Human Genetics, Dublin, Ireland.