Stability and kinetics of c-MYC promoter G-quadruplexes studied by single-molecule manipulation

J Am Chem Soc. 2015 Feb 25;137(7):2424-7. doi: 10.1021/ja511680u. Epub 2015 Feb 13.

Abstract

A DNA G-quadruplex (G4) formed at the oncogene c-MYC promoter region functions as a gene silencer. Due to its high stability at physiological K(+) concentrations, its thermodynamics and kinetic properties have not been characterized in physiological solution conditions. In this work, we investigated the unfolding and folding transitions of single c-MYC G4 and several of its truncated or point mutants at 100 mM KCl concentration under mechanical force. We found that the wild type could fold into multiple species, and the major specie has a slow unfolding rate of (1.4 ± 1.0) × 10(-6) s(-1). The force-dependent thermodynamics and kinetic properties of the major specie were obtained by studying a truncated mutant, Myc2345, that contains the G-tracts 2, 3, 4, and 5. As the c-MYC G4 is a prototype of many other intermolecular parallel-stranded G4's, our results provide important insights into the stability of a broad class of promoter G4's which also play a role in transcription regulation and are potential anticancer targets.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Biomechanical Phenomena / drug effects
  • Dose-Response Relationship, Drug
  • G-Quadruplexes* / drug effects
  • Genes, myc / genetics*
  • Kinetics
  • Mutation
  • Potassium Chloride / pharmacology
  • Promoter Regions, Genetic / drug effects
  • Promoter Regions, Genetic / genetics*
  • Thermodynamics

Substances

  • Potassium Chloride