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Title: Investigation of mechanisms of DNA methylation maintenance regulation in pluripotency
Author: Hay, David James
ISNI:       0000 0004 8510 377X
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2020
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The early stages of development are characterised by profound transcriptional and epigenetic reprogramming. The most striking example is the global loss of DNA methylation patterns on the maternal and paternal genomes that is precipitated upon formation of the zygote. This is coincident with the establishment of the inner-cell mass, a pluripotent population of cells that give rise to the embryo. The necessity to reset the epigenetic landscape to attain this state however requires further investigation, as are the mechanisms that lead to these changes. Using a mouse embryonic stem culture media, referred to as 2i, that models this developmental period, I set out to uncover mechanisms that may contribute to the remodelling of DNA methylation. In contrast to traditional serum-based mouse embryonic stem cell culture media, 2i maintains DNA methylation at a significantly lower level. The use of 2i and serum cultures therefore provided a means to investigate the regulation of the different DNA methylation landscapes. The overarching focus has been to understand how the mediators of DNA methylation, the DNA methyltransferases, are regulated at the level of the protein in response to the stimuli of the media. The differential makeup of the protein complexes of Dnmt1, referred to as the main DNA methyltransferase, and of its obligate co-factor, Uhrf1, were characterised in a proteomic screen. This led to the identification of several candidates and their potential roles in mediating DNA methylation have been postulated to build a case for future investigation. Furthermore, the influence of citrullination, a relatively understudied post-translational modification detected throughout early development, was also investigated in these model culture systems.
Supervisor: Meehan, Richard ; Christophorou, Maria Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
Keywords: DNA methylation ; mouse model ; stem cell ; DNA methyltransferases ; Dnmt1 ; citrullination