CAREER: Uncovering Mechanisms of DNA-protein Crosslink Repair

及时准确的DNA复制对所有真核生物的生长至关重要，但复制过程受到DNA损伤（如DNA-蛋白质交联）的阻碍。持续的dna -蛋白质交联阻止了染色体的分离和向子细胞的传递；因此，去除dna -蛋白交联对细胞存活至关重要。该项目的总体目标是确定细胞如何选择不同的细胞途径来绕过或修复dna -蛋白质交联。结合生物化学、分子生物学和酵母遗传学的多学科策略将被用来研究细胞如何标记dna -蛋白质交联以进行蛋白质水解降解。此外，该项目将吸引生活在农村地区的不同群体的学生，为他们提供科学研究方面的培训、技能和经验，并促进他们参与STEM职业。该研究项目旨在阐明选择性靶向和去除DNA-蛋白交联的生化和分子机制，这些交联在物理上阻碍DNA复制、基因转录和染色质重塑。多种非冗余途径协同修复或绕过dna -蛋白质交联。SPRTN是致力于dna -蛋白交联修复的哺乳动物蛋白酶的创始成员，此后在多个物种中发现了几个旁同源物和同源物，强调了dna -蛋白交联修复的保守性。考虑到可以共价捕获在DNA上的大量蛋白质，SPRTN如何在染色质相关蛋白拥挤的环境中选择性地靶向它们以去除它们仍然未知。目的1将研究针对dna -蛋白质交联的统一机制，无论其大小和化学成分如何。目的2将研究SPRTN蛋白水解活性如何下调以防止染色质相关蛋白的异常破坏。目的3是一种遗传学方法来鉴定出芽酵母中dna -蛋白交联修复的其他重要因素。最后一个目标是让来自农村学校的高中生在自己的教室里进行筛选实验，从而为他们提供STEM职业发展和指导机会。该项目由生物科学理事会分子和细胞生物科学部的遗传机制计划和促进竞争性研究的既定计划（EPSCoR）共同资助。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。

Timely and accurate duplication of DNA is essential for the growth of all eukaryotic organisms, but the replication process is subject to obstruction by DNA damage such as DNA-protein crosslinks. Sustained DNA-protein crosslinks prevent segregation and transmission of chromosomes to daughter cells; hence, removal of DNA-protein crosslinks is critically important for cell survival. The overarching goal of this project is to determine how cells choose distinct cellular pathways to bypass or repair DNA-protein crosslinks. A multidisciplinary strategy combining biochemistry, molecular biology and yeast genetics will be employed to investigate how cells mark DNA-protein crosslinks for proteolytic degradation. Additionally, the project will engage a diverse group of students living in rural areas to provide training, skills, and experiences in scientific research and promote their participation in STEM careers.The research project aims to elucidate the biochemical and molecular mechanisms underlying selective targeting and removal of DNA-protein crosslinks that physically impede DNA replication, gene transcription and chromatin remodeling. Multiple, non-redundant pathways cooperate to repair or bypass DNA-protein crosslinks. SPRTN is the founding member of mammalian proteases dedicated to DNA-protein crosslink repair and several paralogues and orthologues have since been identified in species across multiple kingdoms, underscoring the conserved nature of DNA-protein crosslink repair. Given the multitude of proteins that can be covalently trapped on DNA, how SPRTN selectively targets them for removal in a crowded landscape of chromatin-associated proteins remains unknown. Aim 1 will investigate a unifying mechanism to target DNA-protein crosslinks, irrespective of their size and chemical composition. Aim 2 will investigate how SPRTN proteolytic activity is down-regulated to prevent aberrant destruction of chromatin-associated proteins. Aim 3 is a genetic approach to identify additional factors important for DNA-protein crosslink repair in the budding yeast Saccharomyces cerevisiae. The last goal involves high school students from rural schools performing screening experiments in their own classrooms, and thus provides them with STEM career development and mentoring opportunities.This project is jointly funded by the Genetic Mechanisms program in the Molecular and Cellular Biosciences Division of the Biological Sciences Directorate and the Established Program to Stimulate Competitive Research (EPSCoR).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.