Univ. Bordeaux, INSERM U1212, CNRS UMR 5320, ARNA Laboratory, IECB, F-33607 Pessac, France.
Univ. Toulouse, IPBS, CNRS UMR 5089, 205 route de Narbonne, 31077 Toulouse, France.
Biochim Biophys Acta Gen Subj. 2017 May;1861(5 Pt B):1382-1388. doi: 10.1016/j.bbagen.2017.01.006. Epub 2017 Jan 6.
G-quadruplexes (G4) are RNA and DNA secondary structures formed by the stacking of guanine quartets in guanine rich sequences. Quadruplex-prone motifs may be found in key genomic regions such as telomeres, ribosomal DNA, transcriptional activators and regulators or oncogene promoters. A number of proteins involved in various biological processes are able to interact with G4s. Among them, proteins dedicated to nucleic acids unwinding such as WRN, BLM, FANCJ or PIF1, can unfold G4 structures. Mutations of these helicases are linked to genome instability and to increases in cancer risks. Here, we present a high-throughput fluorescence-based reliable, inexpensive and fast assay to study G4/RHAU interaction. RHAU is an RNA helicase known as the major source of G4 resolution in HeLa cells. Our assay allows to monitor the unfolding properties of RHAU towards DNA and RNA quadruplexes in parallel and to screen for the optimal conditions for its activity. This article is part of a Special Issue entitled "G-quadruplex" Guest Editor: Dr. Concetta Giancola and Dr. Daniela Montesarchio.
四链体（G4）是由富含鸟嘌呤的序列中鸟嘌呤四联体堆叠形成的 RNA 和 DNA 二级结构。四链体倾向基序可能存在于关键基因组区域，如端粒、核糖体 DNA、转录激活剂和调节剂或癌基因启动子。许多参与各种生物过程的蛋白质能够与 G4 相互作用。其中，专门用于解旋核酸的蛋白质，如 WRN、BLM、FANCJ 或 PIF1，可以展开 G4 结构。这些解旋酶的突变与基因组不稳定性和癌症风险增加有关。在这里，我们提出了一种基于荧光的高通量、可靠、廉价和快速的测定法，用于研究 G4/RHAU 相互作用。RHAU 是一种 RNA 解旋酶，被称为 HeLa 细胞中 G4 分辨率的主要来源。我们的测定法允许平行监测 RHAU 对 DNA 和 RNA 四链体的展开特性，并筛选其活性的最佳条件。本文是题为“G-四链体”的特刊的一部分，客座编辑：Concetta Giancola 博士和 Daniela Montesarchio 博士。
