[A photoreactive analogue of 2'3'-dideoxyuridine 5'-triphosphate: preparation and use for photoaffinity modification of human replication protein A]

Bioorg Khim. 2000 Feb;26(2):151-5.
[Article in Russian]


A new reagent for photoaffinity modification of biopolymers, 5-[E-N-(2-nitro-5-azidobenzoyl)-3-amino-1-propen-1-yl]-2',3'-dideoxyuridine 5'-triphosphate (NAB-ddUTP), was synthesized. Like a similar derivative of 2'-deoxyuridine 5'-triphosphate (NAB-dUTP), it was shown to be able to effectively substitute for dTTP in the synthesis of DNA catalyzed by eukaryotic DNA polymerase beta and to terminate DNA synthesis. A 5'-32P-labeled primer with a photoreactive group at the 3'-terminus was derived from NAB-ddUTP and used for photoaffinity labeling of the human replication protein A (RPA). The covalent attachment of RPA p32 and p70 subunits to the labeled primers was demonstrated. NAB-ddUTP is a promising tool for studying the interaction of proteins of the replicative complex with NA in cellular extracts and living cells during the termination of DNA synthesis.

Publication types

  • English Abstract

MeSH terms

  • DNA-Binding Proteins / chemistry*
  • Dideoxynucleotides
  • Humans
  • Photoaffinity Labels
  • Replication Protein A
  • Uracil Nucleotides / chemistry*


  • DNA-Binding Proteins
  • Dideoxynucleotides
  • Photoaffinity Labels
  • RPA1 protein, human
  • Replication Protein A
  • Uracil Nucleotides
  • 2',3'-dideoxyuridine-5'-triphosphate