A DNA polymerase stop assay for G-quadruplex-interactive compounds.
We have developed and characterized an assay for G-quadruplex-interactive compounds that makes use of the fact that G-rich DNA templates present obstacles to DNA synthesis by DNA polymerases. Using Taq DNA polymerase and the G-quadruplex binding 2, 6-diamidoanthraquinone BSU-1051, we find that BSU-1051 leads to enhanced arrest of DNA synthesis in the presence of K+by stabilizing an intramolecular G-quadruplex structure formed by four repeats of either TTGGGG or TTAGGG in the template strand. The data provide additional evidence that BSU-1051 modulates telomerase activity by stabilization of telomeric G-quadruplex DNA and point to a polymerase arrest assay as a sensitive method for screening for G-quadruplex-interactive agents with potential clinical utility.
Han
H
H
Program in Molecular Biology, Division of Medicinal Chemistry and Drug Dynamics Institute, College of Pharmacy, The University of Texas at Austin, Austin, TX 78712, USA. m-salazar@mail.utexas.edu
Hurley
L H
LH
Salazar
M
M
D000880:Anthraquinones / Q000494:pharmacology*; D002414:Cations, Monovalent / Q000494:pharmacology; D004247:DNA / Q000096:biosynthesis*; D054856:G-Quadruplexes; D006150:Guanine Nucleotides / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D011188:Potassium / Q000494:pharmacology; D019914:Taq Polymerase / Q000378:metabolism*; D019098:Telomerase / Q000187:drug effects*
D000880:Anthraquinones; C117697:BSU 1051; D002414:Cations, Monovalent; D006150:Guanine Nucleotides; D004247:DNA; D019914:Taq Polymerase; D019098:Telomerase; D011188:Potassium