Uranyl photofootprinting

Methods Mol Biol. 2009:543:87-96. doi: 10.1007/978-1-60327-015-1_7.

Abstract

The uranyl-(VI) cation (UO(2) (2+)) forms strong complexes with accessible phosphates of nucleic acid (DNA and RNA) backbones. Upon excitation with long wavelength ultraviolet light (lambda = 300-420 nm), uranyl ions bound to backbone phosphates oxidize proximal sugars and induce nucleic acid backbone cleavage. Thus the uranyl(VI) ion functions as a very specific and efficient photochemical probe for identifying ligand(protein)-phosphate contacts in nucleic acid complexes as well as potential (high affinity) cation (e.g., Mg(2+))-binding sites in folded nucleic acids. Finally, the cleavage modulation of duplex DNA reflects helix conformation in terms of minor groove width, due to preferential affinity/oxidation efficiency for such regions of the DNA helix.

MeSH terms

  • Animals
  • Base Sequence
  • Cattle
  • DNA Footprinting / methods*
  • Deoxyribonuclease I / metabolism
  • Light*
  • Molecular Sequence Data
  • Operator Regions, Genetic
  • Repressor Proteins / metabolism
  • Uranium Compounds / chemistry*
  • Viral Regulatory and Accessory Proteins / metabolism

Substances

  • Repressor Proteins
  • Uranium Compounds
  • Viral Regulatory and Accessory Proteins
  • phage repressor proteins
  • Deoxyribonuclease I