M. Protic, S. Hirschfeld, A. P. Tsang, M. Wagner, K. Dixon, A. Levine
{"title":"一种新的损伤特异性DNA结合蛋白的诱导与灵长类细胞中DNA修复的增强相关。","authors":"M. Protic, S. Hirschfeld, A. P. Tsang, M. Wagner, K. Dixon, A. Levine","doi":"10.1201/9781003075387-2","DOIUrl":null,"url":null,"abstract":"Pretreatment of mammalian cell with DNA-damaging agents, such as UV light or mitomycin C, but not the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA), results in the enhanced repair of subsequently transfected UV-damaged expression vectors. To determine the cellular factors that are responsible for this enhancement, we have used a modified gel retardation assay to detect the proteins that interact with damaged DNA. We have identified a constitutive DNA binding protein in extracts from primate cells that has a high affinity for UV-irradiated double-stranded DNA. Cells pretreated with UV light, mitomycin C, or aphidicolin, but not TPA or serum starvation, have higher levels of this damage-specific DNA binding (DDB) protein. These results suggest that the signal for induction of DDB protein can either be damage to the DNA or interference with cellular DNA replication. The induction of DDB protein varies among primate cells with different phenotypes: (1) virus-transformed repair-proficient cells have partially or fully lost the ability to induce DDB protein above constitutive levels; (2) primary cells from repair-deficient xeroderma pigmentosum (XP) group C, and transformed XP groups A and D, show constitutive DDB protein, but do not show induced levels of this protein 48 h after UV; and (3) primary and transformed repair-deficient cells from one XP E patient are lacking both the constitutive and the induced DDB activity. The correlation between the induction of the DDB protein and the enhanced repair of UV-damaged expression vectors implies the involvement of the DDB protein in this inducible cellular response.","PeriodicalId":77750,"journal":{"name":"Molecular toxicology","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"1989-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"18","resultStr":"{\"title\":\"Induction of a novel damage-specific DNA binding protein correlates with enhanced DNA repair in primate cells.\",\"authors\":\"M. Protic, S. Hirschfeld, A. P. Tsang, M. Wagner, K. Dixon, A. 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The induction of DDB protein varies among primate cells with different phenotypes: (1) virus-transformed repair-proficient cells have partially or fully lost the ability to induce DDB protein above constitutive levels; (2) primary cells from repair-deficient xeroderma pigmentosum (XP) group C, and transformed XP groups A and D, show constitutive DDB protein, but do not show induced levels of this protein 48 h after UV; and (3) primary and transformed repair-deficient cells from one XP E patient are lacking both the constitutive and the induced DDB activity. The correlation between the induction of the DDB protein and the enhanced repair of UV-damaged expression vectors implies the involvement of the DDB protein in this inducible cellular response.\",\"PeriodicalId\":77750,\"journal\":{\"name\":\"Molecular toxicology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1989-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"18\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Molecular toxicology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1201/9781003075387-2\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular toxicology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1201/9781003075387-2","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 18
摘要
用dna损伤剂预处理哺乳动物细胞,如紫外线或丝裂霉素C,而不是肿瘤启动子12- o -tetradecanoyl- phorboll -13-acetate (TPA),结果增强了随后转染的紫外线损伤表达载体的修复。为了确定导致这种增强的细胞因素,我们使用了一种改进的凝胶延迟试验来检测与受损DNA相互作用的蛋白质。我们已经从灵长类细胞的提取物中鉴定出一种对紫外线照射的双链DNA具有高亲和力的DNA结合蛋白。用紫外光、丝裂霉素C或阿霉素预处理的细胞,而不是TPA或血清饥饿处理的细胞,具有更高水平的这种损伤特异性DNA结合(DDB)蛋白。这些结果表明,诱导DDB蛋白的信号可能是DNA损伤或干扰细胞DNA复制。在不同表型的灵长类动物细胞中,DDB蛋白的诱导存在差异:(1)病毒转化的修复熟练细胞部分或完全丧失了诱导DDB蛋白高于组成水平的能力;(2)修复缺陷性着色性干皮病(XP) C组和转化后的XP A、D组原代细胞均可见组成性DDB蛋白,但在紫外线照射48 h后未显示该蛋白的诱导水平;(3)来自一名XP E患者的原代和转化修复缺陷细胞既缺乏构成型活性,也缺乏诱导型活性。DDB蛋白的诱导与uv损伤表达载体的增强修复之间的相关性表明DDB蛋白参与了这种可诱导的细胞反应。
Induction of a novel damage-specific DNA binding protein correlates with enhanced DNA repair in primate cells.
Pretreatment of mammalian cell with DNA-damaging agents, such as UV light or mitomycin C, but not the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA), results in the enhanced repair of subsequently transfected UV-damaged expression vectors. To determine the cellular factors that are responsible for this enhancement, we have used a modified gel retardation assay to detect the proteins that interact with damaged DNA. We have identified a constitutive DNA binding protein in extracts from primate cells that has a high affinity for UV-irradiated double-stranded DNA. Cells pretreated with UV light, mitomycin C, or aphidicolin, but not TPA or serum starvation, have higher levels of this damage-specific DNA binding (DDB) protein. These results suggest that the signal for induction of DDB protein can either be damage to the DNA or interference with cellular DNA replication. The induction of DDB protein varies among primate cells with different phenotypes: (1) virus-transformed repair-proficient cells have partially or fully lost the ability to induce DDB protein above constitutive levels; (2) primary cells from repair-deficient xeroderma pigmentosum (XP) group C, and transformed XP groups A and D, show constitutive DDB protein, but do not show induced levels of this protein 48 h after UV; and (3) primary and transformed repair-deficient cells from one XP E patient are lacking both the constitutive and the induced DDB activity. The correlation between the induction of the DDB protein and the enhanced repair of UV-damaged expression vectors implies the involvement of the DDB protein in this inducible cellular response.