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• W2910450861 abstract "DNA replication is one of the most fundamental processes in every living organism,required for the propagation of a cells (epi)genetic information. To ensure the error-freeduplication and transmission of all genetic material exactly once per cell cycle, DNAreplication follows a tightly controlled spatio-temporal program which is conserved acrossmany species. Especially in higher eukaryotic organisms, however, the regulation of thisprogram needs to be dynamic and flexible enough, in order to allow the coordination withother DNA-dependent processes like transcription and DNA repair. Furthermore, earlystudies already showed that the replication timing program undergoes remarkable changesduring cellular development that can be controlled on multiple levels. During the earlyembryonic stages of Drosophila and Xenopus, for example, DNA duplication is achieved inthe order of minutes and depends on the availability of maternal factors that allow rapidDNA synthesis in the absence of transcription, a specialized cell cycle organizationcharacterized by the lack of gap phases which allows cells to oscillate between S-phaseand cell division, as well as differences in the regulation of origin licensing and activationevents. More recently, it was shown that embryonic stem cells of mice and humans are alsosubject to massive rearrangements in their DNA replication timing program duringdevelopment that can affect as much as 50 % of the whole genome.In the course of this work, I took a closer look at these developmental differences inembryonic stem cells of the mouse (mESCs) and analyzed how they manifest in situ. Withthe use of classical nucleotide pulse-chase experiments, I performed a detailedcharacterization of the DNA replication program of mES cells, which was subsequentlyconfirmed in vivo. I observed remarkable differences in the replication timing ofpericentromeric heterochromatin, which may be explained on the basis of differences inchromatin organization of pluripotent cells.With the use of 3-dimensional fluorescence in situ hybridization (3D-FISH), I analyzed thespecific replication timing of three major chromosomal tandem repeat elements, i.e. minorand major satellite repeats and telomeres, and identified the Y-chromosome as the laststructure to be replicated during S-phase in male cells. I could further show that itsduplication occurs in a synchronous manner, similar to that of the inactive X-chromosomeof female cells, which suggests a distinct mode of replication that may be specific to thesetwo inactive chromosomes.Using a combination of single molecule and super-resolution microscopy techniques I wasable to characterize important molecular parameters of the embryonic stem cell replicon,which allowed me to compare the conservation of this crucial functional unit of DNAreplication, with that of somatic cells published in a recent study. These data could indicatefurther developmental differences in the organization of DNA replication, based onmechanisms that might be conserved between mammalian species, frogs and even flies.Last but not least, I analyzed the effect of the loss of DNA methylation on DNA replication.While the lack of this important base modification did not interfere with the globalprogression of the DNA replication machinery, the results show that DNA methylation couldbe important for the control of DNA helix stability and might have the ability to modulateDNA-dependent metabolic processes on the molecular level." @default.
• W2910450861 created "2019-01-25" @default.
• W2910450861 creator A5032973970 @default.
• W2910450861 date "2018-11-19" @default.
• W2910450861 modified "2023-09-23" @default.
• W2910450861 title "DNA replication dynamics in embryonic stem cells" @default.
• W2910450861 hasPublicationYear "2018" @default.
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