Interaction of nucleotide excision repair proteins with DNA containing bulky lesion and apurinic/apyrimidinic site.
The interaction of nucleotide excision repair (NER) proteins (XPC-HR23b, RPA, and XPA) with 48-mer DNA duplexes containing the bulky lesion-mimicking fluorescein-substituted derivative of dUMP (5-{3-[6-(carboxyamidofluoresceinyl)amidocapromoyl]allyl}-2'-deoxyuridine-5'-monophosphate) in a cluster with a lesion of another type (apurinic/apyrimidinic (AP) site) has been studied. It is shown that XPC-HR23b is modified to a greater extent by the DNA duplex containing an AP site opposite nucleotide adjacent to the fluorescein residue than by DNA containing an AP site shifted to the 3'- or 5'-end of the DNA strand. The efficiency of XPA modification by DNA duplexes containing both AP site and fluorescein residue is higher than that by DNA lacking the bulky lesion; the modification pattern in this case depends on the AP site position. In accordance with its major function, RPA interacts more efficiently with single-stranded DNA than with DNA duplexes, including those bearing bulky lesions. The observed interaction between the proteins involved in nucleotide excision repair and DNA structures containing a bulky lesion processed by NER and the AP site repaired via base excision repair may be significant for both these repair pathways in cells and requires the specific sequence of repair of clustered DNA lesions.
Institute of Chemical Biology and Fundamental Medicine, pr. Lavrentieva 8, 630090 Novosibirsk, Russia.
L V
LV
Skosareva
N A
NA
Lebedeva
N I
NI
Rechkunova
E A
EA
Maltseva
P E
PE
Pestryakov
O I
OI
Lavrik
D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004260:DNA Repair; D004277:DNA, Single-Stranded / Q000378:metabolism; D003856:Deoxyuracil Nucleotides / Q000737:chemistry / Q000378:metabolism; D019281:Dimerization; D005452:Fluoresceins / Q000737:chemistry; D006801:Humans; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D051716:Replication Protein A / Q000235:genetics / Q000378:metabolism*; D051760:Xeroderma Pigmentosum Group A Protein / Q000235:genetics / Q000378:metabolism*
D004277:DNA, Single-Stranded; D003856:Deoxyuracil Nucleotides; D005452:Fluoresceins; C497167:RPA1 protein, human; D011994:Recombinant Proteins; D051716:Replication Protein A; C497314:XPA protein, human; D051760:Xeroderma Pigmentosum Group A Protein; D004247:DNA; C007267:2'-deoxyuridylic acid