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，可去除受损DNA中的大块和/或螺旋扭曲病变。转录偶联修复（TCR）和全球基因组修复（GGR）是构成NER的两条途径。TCR是由DNA损伤位点的RNA聚合酶II （RNAPII）的停滞触发的，并致力于快速去除活性转录基因转录链中的病变。真核细胞的TCR是一个复杂的过程，其潜在的分子机制在很大程度上尚未解决。在这个项目中，多种方法，包括酵母遗传学和蛋白质组学，将被用来解决长期存在的关于TCR是如何调节的问题。这项关于基本DNA修复途径的研究将为在面对持续暴露于DNA损伤的情况下维持基因组稳定性的生物过程提供新的见解。该项目还包括实质性的教育组成部分，包括对研究生和本科生的培训，以及对当地学校学生的STEM推广。该项目有三个目标。第一部分将讨论由Rpb4和Rpb7蛋白组成的RNAPII柄如何调节TCR。Rpb4和Rpb7可以抑制或促进TCR，并直接与许多尚未确定的蛋白相互作用。鉴定Rpb4和rpb7相互作用蛋白，并分析其在TCR中的功能。第二个目标将研究TCR促进因子（包括Rad26、Rpb9、Sen1和Elf1）如何相互作用，如何与TCR抑制因子Spt4/Spt5以及RNAPII相互作用，以调节转录病变旁路和TCR。最后，第三个目标将研究TFIIH亚基Tfb1的Pleckstrin同源结构域（PHD）在TCR和GGR中的作用。总之，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。这项研究由生物科学理事会分子和细胞生物科学部的遗传机制项目资助。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值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.