Institute of Molecular Medicine, National Tsing-Hua University, Hsinchu 30013, Taiwan.
Toxicol Sci. 2011 Aug;122(2):339-48. doi: 10.1093/toxsci/kfr107. Epub 2011 May 10.
Nucleotide excision repair (NER) consists of a sequence of events including DNA damage recognition, excision of the damage containing oligonucleotide, gap filling, and ligation. We found that gap filling during the repair of ultraviolet (UV)C-induced DNA lesions was inhibited by various compounds, e.g., amoxicillin, and mixtures, e.g., propolis, the materials that could induce oxidative DNA damage in serum-supplemented cell cultures. Such inhibitory effect was also demonstrated by the immunostaining experiment and host cell reactivation assay. In this study, we link the repair of oxidative DNA damage with the inhibition of gap filling. Our experimental evidence includes the following: (1) induction of oxidative DNA damage and inhibition of gap filling were quantitatively correlated; (2) although the repair of UV-induced DNA damage was delayed in the presence of propolis, the repair of propolis-induced oxidative DNA damage proceeded regardless of preexposure to UV radiation; (3) inhibition of gap filling by propolis was absent in base excision repair (BER)-deficient cells; (4) suppression of propolis-induced oxidative DNA damage by β-carotene abolished the inhibition of gap filling; and (5) inhibition of gap filling was also found with typical BER-inducing agents such as hydrogen peroxide, menadione, and methyl methanesulfonate. We propose that competition may occur between NER and BER, which results in delay of gap filling. Our study reveals the dominancy of BER over NER.
核苷酸切除修复(NER)包括一系列事件,包括 DNA 损伤识别、损伤寡核苷酸的切除、缺口填充和连接。我们发现,在修复紫外线(UV)C 诱导的 DNA 损伤期间,缺口填充被各种化合物抑制,例如阿莫西林,以及混合物,例如蜂胶,这些物质可以在含有血清的细胞培养物中诱导氧化 DNA 损伤。免疫染色实验和宿主细胞复活实验也证明了这种抑制作用。在这项研究中,我们将氧化 DNA 损伤的修复与缺口填充的抑制联系起来。我们的实验证据包括:(1)氧化 DNA 损伤的诱导与缺口填充的抑制呈定量相关;(2)尽管在蜂胶存在的情况下,UV 诱导的 DNA 损伤的修复被延迟,但蜂胶诱导的氧化 DNA 损伤的修复仍然进行,而不管是否预先暴露于 UV 辐射;(3)在碱基切除修复(BER)缺陷细胞中,蜂胶对缺口填充的抑制作用缺失;(4)β-胡萝卜素抑制蜂胶诱导的氧化 DNA 损伤消除了对缺口填充的抑制;(5)典型的 BER 诱导剂,如过氧化氢、甲萘醌和甲基甲烷磺酸盐,也抑制缺口填充。我们提出,NER 和 BER 之间可能存在竞争,导致缺口填充延迟。我们的研究揭示了 BER 在 NER 中的优势。