Year: 2016
Type: dissertation
Abstract: DNA double-strand breaks (DSBs) pose a threat to cell survival and genomic integrity, and remarkable mechanisms exist to deal with these breaks. A single DSB activates a signalling response that profoundly impacts cell physiology, not least through the engagement of DSB repair pathways and the arrest of cell division. Here I study these processes in the budding yeast Saccharomyces cerevisiae and investigate two central themes to this response. First, I examine how the natural ends of chromosomes, telomeres, are differentiated from DSBs by generating DNA ends with increasing telomeric character. I discover a striking transition in the activity of the telomerase inhibitor Pif1 at these ends and propose that this is the dividing line between DSBs and telomeres. Second, I investigate a phenomenon whereby a DSB increases the mobility of chromosomes within the nucleus. This increase in mobility is dependent on the Mec1 kinase and is proposed to promote repair by homologous recombination. I identify that the Mec1-dependent phosphorylation of Cep3, a kinetochore component, is required to stimulate chromatin mobility following DNA breakage and provide a new model for how a DSB affects the constraints on chromosomes. Unexpectedly, I find that increased mobility is not required for DSB repair and instead propose that Cep3 helps arrest the cell cycle in response to a DSB. Finally, my investigation into Cep3 phosphorylation provides new insight into the role of chromatin remodelers and the histone variant H2A.Z in DSB-induced chromatin mobility.
Source: TSpace (University of Toronto)
Author: Jonathan Strecker
Language: English
Topic: DNA Repair Mechanisms
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2756354967