Nucleotide excision repair of 5-formyluracil in vitro is enhanced by the presence of mismatched bases.
5-Formyluracil (fU) is a major thymine lesion produced by reactive oxygen radicals and photosensitized oxidation. Although this residue is a potentially mutagenic lesion and is removed by several base excision repair enzymes, it is unknown whether fU is the substrate of nucleotide excision repair (NER). Here, we analyzed the binding specificity of XPC-HR23B, which initiates NER, and cell-free NER activity on fU opposite four different bases. The result of the gel mobility shift assay showed that XPC-HR23B binds the fU-containing substrates in the following order: fU:C >> fU:T > fU:G > fU:A. Furthermore, in the presence of XPC-HR23B, the dual incision activity was the same as the order of the binding affinity of XPC-HR23B to fU. Therefore, it is concluded that even fU, regarded as a shape mimic of thymine, can be recognized as a substrate of NER incision, and the efficiency depends on instability of the base pair.
Cellular Physiology Laboratory, RIKEN, Japan Science and Technology Corporation, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Katsuhito
K
Kino
Yuichiro
Y
Shimizu
Kaoru
K
Sugasawa
Hiroshi
H
Sugiyama
Fumio
F
Hanaoka
D020137:Base Pair Mismatch; D002474:Cell-Free System / Q000737:chemistry / Q000378:metabolism; D004260:DNA Repair; D045643:DNA Repair Enzymes; D004270:DNA, Circular / Q000737:chemistry / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D006801:Humans; D011485:Protein Binding; D013941:Thymine / Q000737:chemistry / Q000378:metabolism; D014498:Uracil / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000378:metabolism; D014983:Xeroderma Pigmentosum / Q000378:metabolism
D004270:DNA, Circular; D004268:DNA-Binding Proteins; C007777:5-formyluracil; C086985:RAD23A protein, human; C086984:XPC protein, human; D014498:Uracil; D045643:DNA Repair Enzymes; D013941:Thymine