title: G-quadruplexes unfolding by RHAU helicase.
abstract: 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.
authors.affiliation: Univ. Bordeaux, INSERM U1212, CNRS UMR 5320, ARNA Laboratory, IECB, F-33607 Pessac, France.
authors.forename: Nassima Meriem
authors.initials: NM
authors.lastname: Gueddouda
authors.affiliation: Univ. Bordeaux, INSERM U1212, CNRS UMR 5320, ARNA Laboratory, IECB, F-33607 Pessac, France.
authors.forename: Oscar
authors.initials: O
authors.lastname: Mendoza
authors.affiliation: Univ. Toulouse, IPBS, CNRS UMR 5089, 205 route de Narbonne, 31077 Toulouse, France.
authors.forename: Dennis
authors.initials: D
authors.lastname: Gomez
authors.affiliation: Univ. Bordeaux, INSERM U1212, CNRS UMR 5320, ARNA Laboratory, IECB, F-33607 Pessac, France. Electronic address: anne.bourdoncle@u-bordeaux.fr.
authors.forename: Anne
authors.initials: A
authors.lastname: Bourdoncle
authors.affiliation: Univ. Bordeaux, INSERM U1212, CNRS UMR 5320, ARNA Laboratory, IECB, F-33607 Pessac, France. Electronic address: jean-louis.mergny@inserm.fr.
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Mergny
mesh_terms: D053487:DEAD-box RNA Helicases / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D054856:G-Quadruplexes; D057166:High-Throughput Screening Assays; D006801:Humans; D009691:Nucleic Acid Denaturation; D011188:Potassium / Q000737:chemistry / Q000378:metabolism; D011401:Promoter Regions, Genetic; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D012313:RNA / Q000737:chemistry / Q000378:metabolism*; D020871:RNA Stability; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship; D016615:Telomere / Q000737:chemistry / Q000378:metabolism; D013696:Temperature
chemical_list: C489427:MYC protein, human; D016271:Proto-Oncogene Proteins c-myc; D012313:RNA; D004247:DNA; C466988:DHX36 protein, human; D053487:DEAD-box RNA Helicases; D011188:Potassium
keywords: DHX36; G-quadruplex; G4 ligand; G4R1; Helicase; Helicase assay; RHAU