A fluorescence-based helicase assay: application to the screening of G-quadruplex ligands

Nucleic Acids Research
Oscar MendozaJean-Louis Mergny

Abstract

Helicases, enzymes that unwind DNA or RNA structure, are present in the cell nucleus and in the mitochondrion. Although the majority of the helicases unwind DNA or RNA duplexes, some of these proteins are known to resolve unusual structures such as G-quadruplexes (G4) in vitro. G4 may form stable barrier to the progression of molecular motors tracking on DNA. Monitoring G4 unwinding by these enzymes may reveal the mechanisms of the enzymes and provides information about the stability of these structures. In the experiments presented herein, we developed a reliable, inexpensive and rapid fluorescence-based technique to monitor the activity of G4 helicases in real time in a 96-well plate format. This system was used to screen a series of G4 structures and G4 binders for their effect on the Pif1 enzyme, a 5' to 3' DNA helicase. This simple assay should be adaptable to analysis of other helicases and G4 structures.

References

Apr 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·R F MacayaJ Feigon
Jul 5, 2001·Proceedings of the National Academy of Sciences of the United States of America·C SchaffitzelA Plückthun
Nov 1, 2002·Nucleic Acids Research·Anh Tuân Phan, Jean-Louis Mergny
Jul 15, 2004·Journal of the American Chemical Society·Anh Tuân PhanDinshaw J Patel
Aug 27, 2005·Nature·Jean-Baptiste BouléVirginia A Zakian
Aug 29, 2006·Nucleic Acids Research·Jean-Baptiste Boulé, Virginia A Zakian
Jan 31, 2007·Journal of the American Chemical Society·Anne De CianDavid Monchaud
Aug 28, 2007·Nucleic Acids Research·Jean-Baptiste Boulé, Virginia A Zakian
May 30, 2009·Bioinformatics·Oliver StegleJulian Huppert Leon
Jul 1, 2009·Methods in Molecular Biology·Jeong-Hoon Kim, Yeon-Soo Seo
Feb 2, 2010·Biochemical and Biophysical Research Communications·Hidenori TaniNaohiro Noda
Mar 19, 2011·Chemistry : a European Journal·Hélène BertrandMarie-Paule Teulade-Fichou
Mar 2, 2012·Journal of the American Chemical Society·Samir AmraneAnh Tuân Phan
Jul 14, 2012·Organic & Biomolecular Chemistry·Sebastian MüllerShankar Balasubramanian
Feb 21, 2013·Nature Chemistry·Giulia BiffiShankar Balasubramanian
May 10, 2013·Nature·Katrin PaeschkeVirginia A Zakian
Aug 13, 2013·The Journal of Biological Chemistry·Sanjay Kumar BhartiRobert M Brosh
Jun 25, 2014·Proceedings of the National Academy of Sciences of the United States of America·Giang Huong NguyenCurtis C Harris
Nov 14, 2014·Biochemistry·Ramanagouda Ramanagoudr-BhojappaKevin D Raney

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Citations

Feb 18, 2016·Nucleic Acids Research·Oscar MendozaJean-Louis Mergny
Feb 4, 2016·Journal of Medicinal Chemistry·Stephen Neidle
Dec 10, 2015·International Journal of Biological Macromolecules·Fangyuan LiGu Yuan
Feb 9, 2016·Biochimica Et Biophysica Acta·Susanna Cogoi, Luigi E Xodo
Jan 10, 2017·Biochimica Et Biophysica Acta. General Subjects·Nassima Meriem GueddoudaJean-Louis Mergny
Apr 4, 2018·Organic & Biomolecular Chemistry·Josué CarvalhoCarla Cruz
Sep 28, 2018·The Journal of Biological Chemistry·Alicia K ByrdKevin D Raney
Feb 1, 2019·Genes·Leticia Koch Lerner, Julian E Sale
Jul 14, 2016·Chemistry : a European Journal·Aurore De RacheJean-Louis Mergny
Jul 22, 2016·The Journal of Biological Chemistry·Layal SafaCarole Saintomé
Sep 14, 2017·Biochemical Society Transactions·Alicia K Byrd, Kevin D Raney
Apr 6, 2019·Nucleic Acids Research·Imee M A Del MundoKaren M Vasquez
Jan 6, 2017·Current Genetics·Nasim Sabouri
May 31, 2018·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Tara L PetersJonathan H Schatz
Sep 7, 2019·Biochimica Et Biophysica Acta. Molecular Cell Research·Imee M A Del MundoGuliang Wang
Feb 18, 2021·Cell Chemical Biology·Pauline LejaultDavid Monchaud
Dec 31, 2020·International Journal of Molecular Sciences·Corey R NelsonTrushar R Patel
Jan 16, 2021·Chembiochem : a European Journal of Chemical Biology·Mikayel AznauryanVictoria Birkedal
Jul 22, 2020·International Journal of Biological Macromolecules·Zhen-Yong XueLi-Jun Guo
Aug 5, 2021·Journal of the American Chemical Society·Jérémie MitteauxDavid Monchaud

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Methods Mentioned

BETA
electrophoresis
helicase assay
Fluorescence
FRET
fluorescence spectroscopy
helicase
nuclear magnetic resonance

Software Mentioned

cmyc

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