Excellence in Research: Investigation of RECQ1 helicase in DNA transactions upon oxidative stress

每个细胞的特征都是由它的DNA决定的，DNA是由两条缠绕在一起的单链DNA组成的双链螺旋分子。在正常的生命过程中，细胞承受DNA损伤，必须修复以保持遗传完整性。该项目将研究RECQ1，一种分离DNA两条链的DNA解旋酶，是如何帮助修复DNA损伤和维持基因组稳定性的。RecQ解旋酶存在于所有生命领域中，因此这项工作将具有广泛的相关性。这项研究将促进研究生和本科生的职业发展。这项工作将提供获得指导研究课程学分或霍华德大学生物化学和分子生物学系荣誉论文的机会。基本实验室技能和跨学科研究培训的实践培训将为学生在研究生教育以及未来在学术界或工业生物技术领域的科学职业生涯中取得成功做好准备。PI还通过美国大学妇女协会、美国国立卫生研究院有色人种妇女科学协会以及美国国家科学基金会的另一项名为HU-ADVANCE-IT的倡议，扩大妇女在科学领域的参与。本研究的核心假设是，RECQ1作为氧化应激引发的遗传不稳定的屏障，并且RECQ1与氧化损伤染色质的功能关联确保了遗传信息的忠实表达和传播。第一个目标是确定RECQ1在氧化应激下DNA损伤反应的激活和氧化DNA损伤修复过程中复制叉进程中的作用。在第二个目标中，将采用无偏倚的生化和细胞方法来确定RECQ1及其相互作用的蛋白质伙伴的催化活性，使其能够促进氧化DNA损伤的修复。最终目的是测试RECQ1对氧化DNA损伤的转录反应的影响，以及这是否通过与易氧化或形成g -四联体的基因组区域的特异性结合来介导。总的来说，本研究将确定氧化应激下RECQ1解旋酶在复制、修复和转录控制水平上的确切作用。这些知识也将有助于确定不同的RecQ家族成员是否具有功能冗余，或者它们是否具有特殊的功能，这些功能可能在调节细胞中的氧化（和其他基因毒性）应激方面至关重要。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。

The characteristics of each cell are defined by its DNA, a double-stranded helical molecule composed of two intertwined single strands of DNA. During the normal course of life cells sustain DNA damage, which must be repaired to maintain genetic integrity. This project will investigate how RECQ1, a DNA helicase that separates the two strands of DNA, helps repair DNA damage and maintain genome stability. RecQ helicases are present in all kingdoms of life and therefore this work will have broad relevance. This research will promote the careers of both graduate and undergraduate students. The work will provide opportunities to earn course credits for Directed Research or for an Honors thesis in the Department of Biochemistry and Molecular Biology at Howard University. Hands-on training in fundamental laboratory skills and interdisciplinary research training will prepare students for success in graduate education and in their future scientific careers in academia or in industrial biotechnology. The PI also performs outreach to broaden participation of women in science through the American Association of University Women, the NIH Women of Color in Science, and through another NSF initiative called HU-ADVANCE-IT.The central hypothesis of this project is that RECQ1 acts as a barrier against genetic instability triggered by oxidative stress and the functional association of RECQ1 with oxidatively damaged chromatin ensures faithful expression and propagation of genetic information. The first aim is to establish the role of RECQ1 in activation of DNA damage response to oxidative stress and in replication fork progression during repair of oxidative DNA damage. In the second aim, unbiased biochemical and cellular approaches will be employed to determine catalytic activities of RECQ1 and its interacting protein partners that allow it to facilitate repair of oxidative DNA damage. The final aim will test the impact of RECQ1 on transcriptional response to oxidative DNA damage and whether this is mediated through specific binding to genomic regions susceptible to oxidation or form G-quadruplexes. Collectively, this research will ascertain the exact role of RECQ1 helicase under oxidative stress at the levels of replication, repair and transcriptional control. This knowledge will also help determine whether different RecQ family members have functional redundancies or whether they have specialized functions that may be critical in regulating oxidative (and other genotoxic) stress in the cell.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.