Analysis of HPRT and supF Mutations Caused by Pierisin-1, a Guanine Specific ADP-Ribosylating Toxin Derived from the Cabbage Butterfly
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- 作者：Yukari Totsuka ; Masanobu Kawanishi ; Rena Nishigaki ; Kazuki Matsukawa ; Takashi Yagi ; Takeji Takamura-Enya ; Masahiko Watanabe ; Takashi Sugimura ; and Keiji Wakabayashi
- 刊名：Chemical Research in Toxicology
- 出版年：2003
- 出版时间：August 2003
- 年：2003
- 卷：16
- 期：8
- 页码：945 - 952
- 全文大小：156K
- 年卷期：v.16,no.8(August 2003)
- ISSN：1520-5010
文摘
Pierisin-1, an ADP-ribosylating toxin derived from the cabbage butterfly, Pieris rapae, inducesapoptosis in various mammalian cell lines. We recently reported that the target for ADPribosylation by pierisin-1 is the 2'-deoxyguanosine residue in DNA. To examine whetherpierisin-1 would induce mutations in mammalian cell genes, we conducted a mutational analysisfor the hypoxanthine-guanine phosphoribosyltransferase (HPRT) locus in pierisin-1-treatedChinese hamster lung (CHL) cells. N2-(ADP-ribos-1-yl)-2'-deoxyguanosine was detected by the32P-postlabeling method in CHL cells after treatment with pierisin-1 at doses of 2-32 ng/mL;adduct levels were 1.1-12.0 per 106 nucleotides. Pierisin-1 induced mutations in the HPRTgene dose-dependently, and the frequency was 38 times higher than the control, at a dose of32 ng/mL. To confirm that mono(ADP-ribosyl)ated dG itself leads to mutations, the pierisin-1-treated DNA of plasmid pMY189 bearing the supF gene was used for mutational analysis.The mutation frequency of the supF gene treated with 2-8 
g/mL of pierisin-1 was 17-40-fold the control value. Mutation spectrum analysis showed that single base substitutionsdominated in both HPRT and supF genes. Among these, transversions were predominant, andmore than 70% of the base substitutions occurred at G:C base pairs in both genes. The mostfrequent mutations were G:C to C:G, followed by G:C to T:A in HPRT gene, whereas G:C toT:A transversions dominated in the supF gene. Our results indicate that pierisin-1 producedN2-(ADP-ribos-1-yl)-2'-deoxyguanosine and this guanine-adduct could lead to mutations in theHPRT and supF genes. These findings could provide very useful information for understandingthe biological significance of pierisin-1.
g/mL of pierisin-1 was 17-40-fold the control value. Mutation spectrum analysis showed that single base substitutionsdominated in both HPRT and supF genes. Among these, transversions were predominant, andmore than 70% of the base substitutions occurred at G:C base pairs in both genes. The mostfrequent mutations were G:C to C:G, followed by G:C to T:A in HPRT gene, whereas G:C toT:A transversions dominated in the supF gene. Our results indicate that pierisin-1 producedN2-(ADP-ribos-1-yl)-2'-deoxyguanosine and this guanine-adduct could lead to mutations in theHPRT and supF genes. These findings could provide very useful information for understandingthe biological significance of pierisin-1.