Regulation of transcription coupled DNA repair

核苷酸切除修复(NER)是一种保守的DNA修复过程，它移除受损DNA中巨大的和/或螺旋扭曲的损伤。转录偶联修复(TCR)和整体基因组修复(GGR)是构成NER的两条途径。TCR是由RNA聚合酶II(RNAPII)在DNA损伤部位的停滞触发的，致力于快速移除活跃转录基因转录链中的损伤。真核细胞中的TCR是一个复杂的过程，其潜在的分子机制在很大程度上仍未解决。在这个项目中，将使用包括酵母遗传学和蛋白质组学在内的各种方法来解决TCR如何受到监管的长期存在的问题。这项关于基本DNA修复途径的研究将对生物过程产生新的见解，这些过程在面对持续的DNA损伤时保持基因组的稳定性。该项目还包括实质性的教育内容，包括对研究生和本科生的培训以及与当地学校学生的STEM外联活动。该项目有三个目标。第一个将讨论RNAPII茎是如何调节TCR的，它由Rpb4和Rpb7蛋白组成。Rpb4和Rpb7可以抑制或促进TCR，并直接与许多尚未鉴定的蛋白质相互作用。将鉴定Rpb4和Rpb7相互作用的蛋白，并分析它们在TCR中的功能。第二个目标将研究TCR促进剂，包括Rad26，Rpb9，Sen1和Elf1，如何相互作用，与TCR抑制子Spt4/Spt5，以及与RNAPII，以调节转录损伤旁路和TCR。最后，第三个目标将研究Tfb1的Pleckstrin同源结构域(PHD)，TFIIH亚基，在TCR和GGR中的作用。这些研究将促进我们对转录偶联DNA修复的分子机制的理解。这项研究是由生物科学局分子和细胞生物科学部的遗传机制项目资助的。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Nucleotide excision repair (NER) is a conserved DNA repair process that removes bulky and/or helix-distorting lesions in damaged DNA. Transcription coupled repair (TCR) and global genomic repair (GGR) are two pathways that constitute NER. TCR is triggered by stalling of RNA polymerase II (RNAPII) at DNA damage sites and is dedicated to rapid removal of lesions in the transcribed strand of actively transcribed genes. TCR in eukaryotic cells is a complicated process and the underlying molecular mechanisms remain largely unresolved. In this project, a variety of approaches, including yeast genetics and proteomics, will be used to address long-standing questions about how TCR is regulated. This research on a fundamental DNA repair pathway will yield new insights into biological processes that maintain genome stability in the face of constant exposure to DNA damage. The project also includes substantive educational components, including training of graduate and undergraduate students as well as STEM outreach to local school students. The project has three objectives. The first will address how the RNAPII stalk, consisting of Rpb4 and Rpb7 proteins, regulates TCR. Rpb4 and Rpb7 can either repress or facilitate TCR and directly interact with many as-yet-unidentified proteins. Rpb4 and Rpb7-interacting proteins will be identified and their functions in TCR will be analyzed. The second objective will examine how TCR facilitators, including Rad26, Rpb9, Sen1 and Elf1, interplay with each other, with the TCR repressors Spt4/Spt5, and with RNAPII to regulate transcriptional lesion bypass and TCR. Finally, the third objective will investigate the role of the Pleckstrin homology domain (PHD) of Tfb1, a TFIIH subunit, in TCR and GGR. Together these studies will advance our understanding of the molecular mechanisms of transcription coupled DNA repair.This research is funded by the Genetic Mechanisms program in the Division of Molecular and Cellular Biosciences in the Directorate of Biological Sciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.