Department of Biochemistry and Molecular Biology, College of Medicine, Howard University, Washington, DC, United States.
Department of Biochemistry and Molecular Biology, College of Medicine, Howard University, Washington, DC, United States; National Human Genome Center, College of Medicine, Howard University, Washington, DC, United States.
Methods Enzymol. 2022;673:227-249. doi: 10.1016/bs.mie.2022.03.057. Epub 2022 Apr 18.
RecQ helicases participate in a variety of DNA metabolic processes through their multiple biochemical activities. In vitro characterization and cellular studies have suggested that RECQ1 (also known as RECQL or RECQL1) performs its diverse functions through specific interactions with DNA and protein partners. We have taken an unbiased approach to determine the contribution of RECQ1 in genome maintenance and as a putative susceptibility factor in breast cancer. Here, we provide methodology to map the genome-wide binding sites of RECQ1 together with the profiling of RECQ1-dependent transcriptome to investigate its role in gene regulation. The described approach will be helpful to develop a mechanistic framework for elucidating critical functions of RECQ1 and other RecQ homologs in distinct chromatin and biological contexts.
RecQ 解旋酶通过其多种生化活性参与多种 DNA 代谢过程。体外特征分析和细胞研究表明,RECQ1(也称为 RECQL 或 RECQL1)通过与 DNA 和蛋白伴侣的特异性相互作用来发挥其多种功能。我们采用了一种无偏见的方法来确定 RECQ1 在基因组维护中的贡献,以及作为乳腺癌中潜在的易感因素。在这里,我们提供了一种绘制 RECQ1 全基因组结合位点图谱的方法,并对 RECQ1 依赖性转录组进行了分析,以研究其在基因调控中的作用。所描述的方法将有助于为阐明 RECQ1 和其他 RecQ 同源物在不同染色质和生物学环境中的关键功能建立一个机制框架。