Repairing the Sickle Cell mutation. III. Effect of irradiation wavelength on the specificity and type of photoproduct formed by a 3'-terminal psoralen on a third strand directed to the mutant base pair

Nucleic Acids Res. 2003 Aug 15;31(16):4682-8. doi: 10.1093/nar/gkg658.

Abstract

Using a psoralen delivery system mediated by a DNA third strand that binds selectively to linear target duplexes immediately downstream from the Sickle Cell beta-globin gene mutation and the comparable wild-type beta-globin gene sequence, the kinetics of formation and yield of psoralen monoadducts and crosslinks with pyrimidine residues at and near the mutant base pair site and its wild-type counterpart were determined. By exploiting irradiation specificities at 300, 365 and 419 nm, it was possible to evaluate the orientation equilibrium of 3'-linked intercalated psoralen and to develop conditions that lead to preferential formation of each type of photoproduct in both the mutant and wild-type sequences. This makes possible the preparation of each type of photoproduct for use as a substrate for DNA repair. In this way, the base pair change(s) that each generates can be established.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Anemia, Sickle Cell / genetics*
  • Base Pairing / genetics
  • Base Sequence
  • DNA / chemistry
  • DNA / metabolism
  • DNA / radiation effects
  • DNA Adducts / chemistry
  • DNA Adducts / metabolism
  • DNA Repair*
  • Electrophoresis, Polyacrylamide Gel / methods
  • Furocoumarins / chemistry
  • Furocoumarins / metabolism
  • Hemoglobin, Sickle / genetics*
  • Humans
  • Kinetics
  • Nucleic Acid Conformation
  • Oligonucleotides / chemistry
  • Oligonucleotides / genetics
  • Point Mutation*
  • Ultraviolet Rays

Substances

  • DNA Adducts
  • Furocoumarins
  • Hemoglobin, Sickle
  • Oligonucleotides
  • psoralen-DNA adduct
  • DNA