[A photoreactive analogue of 2'3'-dideoxyuridine 5'-triphosphate: preparation and use for photoaffinity modification of human replication protein A].
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.
Kolpashchikov
D M
DM
Novosibirsk Institute of Bioorganic Chemistry, Siberian Division, Russian Academy of Sciences, Novosibirsk, Russia.
Aleksandrova
L A
LA
Zakirova
N F
NF
Khodyreva
S N
SN
Lavrik
O I
OI
D004268:DNA-Binding Proteins / Q000737:chemistry*; D054306:Dideoxynucleotides; D006801:Humans; D019920:Photoaffinity Labels; D051716:Replication Protein A; D014500:Uracil Nucleotides / Q000737:chemistry*
D004268:DNA-Binding Proteins; D054306:Dideoxynucleotides; D019920:Photoaffinity Labels; C497167:RPA1 protein, human; D051716:Replication Protein A; D014500:Uracil Nucleotides; C063034:2',3'-dideoxyuridine-5'-triphosphate