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

卓越研究：氧化应激下 RECQ1 解旋酶在 DNA 处理中的研究

• 批准号: 1832163
• 负责人: Sudha Sharma
• 金额: $ 61.18万
• 依托单位: Howard University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1832163&HistoricalAwards=false
• 关键词: Excellence; Research; Investigation; RECQ1; helicase

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.

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，细胞遭受DNA损伤，必须修复才能保持遗传完整性。这个项目将研究RECQ1，一种将DNA的两条链分开的DNA解旋酶，如何帮助修复DNA损伤和保持基因组的稳定。RECQ解旋酶存在于生命的所有王国中，因此这项工作将具有广泛的相关性。这项研究将对研究生和本科生的职业生涯都有促进作用。这项工作将为霍华德大学生物化学和分子生物学系的指导性研究或荣誉论文提供获得课程学分的机会。基础实验室技能实践培训和跨学科研究培训将为学生在研究生教育和他们未来在学术界或工业生物技术的科学职业生涯中取得成功做好准备。PI还通过美国大学女性协会、NIH有色人种科学女性协会以及另一项名为HU-Advance-It的NSF倡议进行推广，以扩大女性在科学领域的参与。该项目的中心假设是，RECQ1起到了抵御氧化应激引发的遗传不稳定的屏障作用，RECQ1与氧化损伤的染色质的功能联系确保了遗传信息的忠实表达和传播。第一个目的是确定RECQ1在DNA损伤的激活和氧化应激反应中的作用，以及在DNA氧化损伤修复过程中的复制分叉过程。在第二个目标中，将使用无偏见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DNA氧化损伤的修复。最终目的将测试RECQ1对氧化DNA损伤的转录反应的影响，以及这是通过与易氧化的基因组区域的特异性结合或形成G-四链来介导的。总而言之，这项研究将在复制、修复和转录控制水平上确定RECQ1解旋酶在氧化应激下的确切作用。这一知识还将有助于确定不同的RecQ家族成员是否具有功能冗余，或者他们是否具有可能在调节细胞中的氧化(和其他基因毒性)压力方面至关重要的专门功能。该奖项反映了NSF的法定使命，并已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。

项目成果

Genome-wide investigations on regulatory functions of RECQ1 helicase.

• DOI: 10.1016/j.ymeth.2022.02.010
• 发表时间: 2022-08
• 期刊: METHODS
• 影响因子: 4.8
• 作者: Debnath, Subrata;Lu, Xing;Lal, Ashish;Sharma, Sudha
• 通讯作者: Sharma, Sudha