Impact of DNA base modifications on the double helix stability and genome activity

DNA碱基修饰对双螺旋稳定性和基因组活性的影响

• 批准号: 413888330
• 负责人: Professorin Dr. Maria Cristina Cardoso
• 金额: --
• 依托单位: Arbeitsgruppe Cell Biology and Epigenetics
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/413888330?language=en
• 关键词: Impact; DNA; base; modifications; double

DNA modifications in eukaryotic organisms are important epigenetic marks and play a central role in regulation of genome activity and stability. While most research is geared towards proteins recognizing and translating this epigenetic information, we want to investigate how these DNA modifications directly and indirectly affect the stability of the DNA double helix and how that affects basic cellular processes like transcription and replication.Local destabilization of the DNA double helix could affect processes like base flipping to modify or repair bases, open complex formation during transcription initiation or modulate the speed of transcription and thereby affect associated processes like RNA processing or RNA guided epigenetic modifications. We will directly measure the effect of known DNA modifications on DNA double helix stability and on DNA/RNA polymerases and DNA helicases in vitro. We will then increase or decrease the level of base modifications using genetic approaches in somatic as well as isogenic embryonic stem cells and will measure rate of ribonucleotide as well as desoxyribonucleotide polymerization in vivo. We will also measure the rate of DNA unwinding by DNA helicase in vivo by uncoupling the DNA polymerization from the DNA helicase activities within the cellular replisome.

真核生物中的DNA修饰是重要的表观遗传标记，在基因组活性和稳定性的调节中起着核心作用。虽然大多数研究都是针对蛋白质识别和翻译这些表观遗传信息，但我们希望研究这些DNA修饰如何直接和间接影响DNA双螺旋的稳定性，以及如何影响转录和复制等基本细胞过程。DNA双螺旋的局部不稳定可能影响碱基翻转等过程，以修改或修复碱基，在转录起始期间开放复合物形成或调节转录速度，从而影响相关过程，如RNA加工或RNA引导的表观遗传修饰。我们将直接测量已知的DNA修饰对DNA双螺旋稳定性和DNA/RNA聚合酶和DNA解旋酶的影响。然后，我们将增加或减少使用体细胞以及同基因胚胎干细胞中的遗传方法的碱基修饰的水平，并将在体内测量核糖核苷酸以及脱氧核糖核苷酸聚合的速率。我们还将通过将DNA聚合与细胞复制体内的DNA解旋酶活性解偶联来测量体内DNA解旋酶的DNA解旋速率。