Quantitative Determination of DNA Bridging Efficiency of Chromatin Proteins

Methods Mol Biol. 2018:1837:199-209. doi: 10.1007/978-1-4939-8675-0_12.

Abstract

DNA looping is important for genome organization in all domains of life. The basis of DNA loop formation is the bridging of two separate DNA double helices. Detecting DNA bridge formation generally involves the use of complex single-molecule techniques (atomic force microscopy, magnetic, or optical tweezers). Although DNA bridging can be qualitatively described, quantification of DNA bridging and bridging dynamics using these techniques is challenging. Here, we describe a novel biochemical assay capable of not only detecting DNA bridge formation, but also allowing for quantification of DNA bridging efficiency and the effects of physico-chemical conditions on DNA bridge formation.

Keywords: DNA bridging; DNA bridging proteins; DNA looping; DNA-DNA cross-linking; DNA-DNA interactions; Pull-down assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Pairing*
  • Chromatin / chemistry*
  • Chromatin / genetics
  • Chromatin / metabolism*
  • DNA / chemistry*
  • DNA / metabolism*
  • DNA-Binding Proteins
  • Isotope Labeling
  • Nucleic Acid Conformation*

Substances

  • Chromatin
  • DNA-Binding Proteins
  • DNA