Molecular Evidence of the Involvement of the Nucleotide Excision Repair (NER) System in the Repair of the Mono(ADP-Ribosyl)ated DNA Adduct Produced by Pierisin-1, an Apoptosis-Inducing Protein from th

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• 作者：Masanobu Kawanishi ; Kazuki Matsukawa ; Isao Kuraoka ; Takeji Takamura-Enya ; Yukari Totsuka ; Yasuko Matsumoto ; Masahiko Watanabe ; Yue Zou ; Kiyoji Tanaka ; Takashi Sugimura ; Keiji Wakabayashi ; Takashi Yagi
• 刊名：Chemical Research in Toxicology
• 出版年：2007
• 出版时间：April 2007
• 年：2007
• 卷：20
• 期：4
• 页码：694 - 700
• 全文大小：276K
• 年卷期：v.20,no.4(April 2007)
• ISSN：1520-5010

文摘

Pierisin-1 is a potent apoptosis-inducing protein found in the pupal extract of the cabbage white butterfly.Pierisin-1 catalyzes the mono(ADP-ribosyl)ation of the 2'-deoxyguanosine residue and produces a bulkyadduct, N²-(ADP-ribos-1-yl)-2'-deoxyguanosine (N²-ADPR-dG) in DNA. Here, we examined theinvolvement of the nucleotide excision repair (NER) system in the removal of N²-ADPR-dG in Escherichiacoli (E. coli) and human cells. The results of mobility shift gel electrophoresis assays using a 50-meroligodeoxynucleotide containing a single N²-ADPR-dG showed that E. coli UvrAB proteins bound tothe N²-ADPR-dG in vitro. Incubation of the adducted oligodeoxynucleotides with UvrABC resulted inthe incision of the oligonucleotides in vitro. The results of filter binding and gel mobility shift assaysusing human XPA protein showed that XPA bound to DNA containing N²-ADPR-dGs in vitro. Finally,we introduced plasmids containing N²-ADPR-dGs into E. coli and human cells. N²-ADPR-adductedplasmids replicated l0 times and 20 times less efficiently in NER-deficient E. coli and human cells thanin their wild-type counterparts, respectively. More mutations were induced in the plasmid propagated inNER-deficient cells than that in wild-type human cells. These results indicate the involvement of theNER system in the repair of N²-ADPR-dG in both E. coli and human cells.