Rec-A protein-mediated irreversible fixation of an oligodeoxyribonucleotide to specific site in DNA

Mutat Res. 1996 Apr 13;351(2):117-24. doi: 10.1016/0027-5107(95)00212-x.

Abstract

RecA protein can polymerize on an oligodeoxyribonucleotide to form a filament that finds its homologous sequence in double-stranded DNA. When such an oligonucleotide is linked to psoralen, a photoactivatable DNA intercalator, it irreversibly binds to the homologous site in double stranded DNA as a result of psoralen photoadduct formation at thymidines. The relative efficiency of specific vs. nonspecific binding of an oligonucleotide depended upon the ratio of psoralenated oligonucleotide to total DNA. Na+ ions at concentrations greater than 50 mM eliminated specific binding. Under optimal conditions. the probability of binding of an 80-mer oligonucleotide to a specific site was > 10(5) times greater than that of binding to any single nonspecific site. Under the conditions described, RecA-mediated photoadduction was equally efficient with superhelical and linear double-stranded DNA.

Publication types

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

MeSH terms

  • Base Sequence
  • Binding Sites
  • DNA Adducts / biosynthesis*
  • Ficusin / metabolism*
  • Molecular Sequence Data
  • Oligodeoxyribonucleotides / metabolism*
  • Plasmids / metabolism*
  • Rec A Recombinases / metabolism*
  • Sequence Homology, Nucleic Acid

Substances

  • DNA Adducts
  • Oligodeoxyribonucleotides
  • Rec A Recombinases
  • Ficusin