Hastert, Florian D. (2019)
Characterization of the cancer-associated short isoform of Tet1 in DNA replication.
Technische Universität Darmstadt
Dissertation, Erstveröffentlichung
Kurzbeschreibung (Abstract)
Almost all cells of a metazoan organism share the same genetic information but can differ highly in their function and phenotype. The transition from the same genotype to different phenotypes is mostly achieved by epigenetic mechanisms, including DNA base modifications. The most prominent and best studied DNA modification is 5-methylcytosine (5mC), which was implicated in transcriptional repression. The levels of 5mC are maintained on the newly synthesized DNA strand, to ensure its faithful inheritance during mitotic cell division. Oxidation of the 5mC by Tet dioxygenases, to 5-hydroxymethylcytosine (5hmC) and even further, can drastically change the epigenetic information transported by this modification and hence modulate gene expression and the overall phenotype of a cell. However, the exact regulation of Tet proteins and the putative cell cycle dependent inheritance of their oxidative products still is under investigation. In the present work, I therefore characterized the subnuclear distribution of different Tet proteins throughout the cell cycle and especially during S-phase and potential implications for cellular homoeostasis. I identified the recruitment of the short isoform of Tet1 (Tet1s), which is found to be overexpressed in different cancers, to sites of ongoing DNA replication in pericentric heterochromatin, which is rich in 5mC. I furthermore found that Tet1s localization during S-phase and also its catalytic activity is highly dependent on a conserved lysine residue. Moreover, I could show that Tet1 physically interacts with the E3 ubiquitin-ligase Uhrf1 and that the observed localization of Tet1s to sites of ongoing DNA replication in pericentric heterochromatin requires the presence of Uhrf1. Finally, I found that a breast cancer cell line that was shown to overexpress Tet1s, exhibits significantly changed 5mC and 5hmC levels, globally as well as in situ, in comparison to non-transformed breast epithelial cells. In summary, my findings contribute to the understanding of how the epigenetic information of every cell can be diversified by the regulation of Tet protein localization and their catalytic activity.
Typ des Eintrags: | Dissertation | ||||
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Erschienen: | 2019 | ||||
Autor(en): | Hastert, Florian D. | ||||
Art des Eintrags: | Erstveröffentlichung | ||||
Titel: | Characterization of the cancer-associated short isoform of Tet1 in DNA replication | ||||
Sprache: | Englisch | ||||
Referenten: | Cardoso, Prof. Dr. M. Cristina ; Laube, Prof. Dr. Bodo | ||||
Publikationsjahr: | 2019 | ||||
Ort: | Darmstadt | ||||
Datum der mündlichen Prüfung: | 25 Januar 2019 | ||||
URL / URN: | https://tuprints.ulb.tu-darmstadt.de/8420 | ||||
Kurzbeschreibung (Abstract): | Almost all cells of a metazoan organism share the same genetic information but can differ highly in their function and phenotype. The transition from the same genotype to different phenotypes is mostly achieved by epigenetic mechanisms, including DNA base modifications. The most prominent and best studied DNA modification is 5-methylcytosine (5mC), which was implicated in transcriptional repression. The levels of 5mC are maintained on the newly synthesized DNA strand, to ensure its faithful inheritance during mitotic cell division. Oxidation of the 5mC by Tet dioxygenases, to 5-hydroxymethylcytosine (5hmC) and even further, can drastically change the epigenetic information transported by this modification and hence modulate gene expression and the overall phenotype of a cell. However, the exact regulation of Tet proteins and the putative cell cycle dependent inheritance of their oxidative products still is under investigation. In the present work, I therefore characterized the subnuclear distribution of different Tet proteins throughout the cell cycle and especially during S-phase and potential implications for cellular homoeostasis. I identified the recruitment of the short isoform of Tet1 (Tet1s), which is found to be overexpressed in different cancers, to sites of ongoing DNA replication in pericentric heterochromatin, which is rich in 5mC. I furthermore found that Tet1s localization during S-phase and also its catalytic activity is highly dependent on a conserved lysine residue. Moreover, I could show that Tet1 physically interacts with the E3 ubiquitin-ligase Uhrf1 and that the observed localization of Tet1s to sites of ongoing DNA replication in pericentric heterochromatin requires the presence of Uhrf1. Finally, I found that a breast cancer cell line that was shown to overexpress Tet1s, exhibits significantly changed 5mC and 5hmC levels, globally as well as in situ, in comparison to non-transformed breast epithelial cells. In summary, my findings contribute to the understanding of how the epigenetic information of every cell can be diversified by the regulation of Tet protein localization and their catalytic activity. |
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URN: | urn:nbn:de:tuda-tuprints-84201 | ||||
Sachgruppe der Dewey Dezimalklassifikatin (DDC): | 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie | ||||
Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie 10 Fachbereich Biologie > Cell Biology and Epigenetics |
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Hinterlegungsdatum: | 03 Mär 2019 20:55 | ||||
Letzte Änderung: | 03 Mär 2019 20:55 | ||||
PPN: | |||||
Referenten: | Cardoso, Prof. Dr. M. Cristina ; Laube, Prof. Dr. Bodo | ||||
Datum der mündlichen Prüfung / Verteidigung / mdl. Prüfung: | 25 Januar 2019 | ||||
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