具有最小紫外线敏感性的FAECB(细胞生物学自动化实验设施)中国仓鼠卵巢突变体的表型

David B Busch , Deborah White Ziffer , Donna Coleman , Lisa Wills , H Greg McDonough , Nigel J Jones
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引用次数: 2

摘要

细胞生物学自动化实验设施(FAECB)收集了200多株紫外线(UV)敏感突变体中国仓鼠卵巢(CHO)细胞,并对其进行了互补组分配(CG)研究,确定了啮齿动物UV CGs 1-6 (ERCC1-6)和新的啮齿动物XRCC9/FANCG组的代表。收集的10个突变体,包括另外6个来自野生型AA8, 3个CHO ERCC1细胞系u4的紫外线敏感双突变体,以及CHO XRCC1细胞系EM9的紫外线敏感突变体,在这些先前的研究中未被指定或鉴定。这10个突变体中,有8个突变体的紫外线敏感性是亲本系(AA8、EM9或u4)的1.5倍左右,2个突变体的紫外线敏感性是AA8的2倍左右。本研究报道了这10个突变体在紫外线(含和不含咖啡因)、电离辐射、丝裂霉素C (MMC)和甲磺酸乙酯(EMS)敏感性方面的部分特征;熟练掌握DNA修复(非计划DNA合成);和UV-mutability。10个细胞系的表型具有异质性,许多细胞系表现出咖啡因对UDS或uv的敏感性降低,而另一些细胞系则表现出对EMS或MMC的明显敏感性,它们可能在参与核苷酸切除修复、复制后修复、碱基切除修复或重组修复的不同基因上发生突变。作为FAECB收集的一部分,先前分离的突变体已被证明在表征哺乳动物DNA修复过程和克隆人类修复基因方面非常重要,而这些当前的突变体虽然对紫外线不敏感,但仍有可能做出进一步的贡献。
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Phenotype of FAECB (Facility for Automated Experiments in Cell Biology) Chinese hamster ovary mutants with minimal UV-sensitivity

The Facility for Automated Experiments in Cell Biology (FAECB) collection of over 200 lines of ultraviolet (UV)-sensitive mutant Chinese hamster ovary (CHO) cells has previously been studied for complementation group assignment (CG), with representatives of rodent UV CGs 1–6 (ERCC1–6) and the new rodent XRCC9/FANCG group identified. Ten mutants from the collection, including a further six derived from wildtype AA8, three UV-sensitive double-mutants of CHO ERCC1 cell line UV4, and a UV-sensitive mutant of CHO XRCC1 cell line EM9, had not been assigned or characterized in these previous studies. These 10 mutants include 8 with approximately 1.5-fold the UV-sensitivity of the parental line (AA8, EM9, or UV4), and 2 with about 2-fold the UV-sensitivity of AA8. The present study reports the partial characterization of these 10 mutants in terms of sensitivity to UV (with and without caffeine), ionizing radiation, mitomycin C (MMC) and ethyl methanesulfonate (EMS); proficiency in DNA repair (unscheduled DNA synthesis (UDS)); and UV-mutability. The phenotypes of the 10 cell lines were heterogeneous, a number showed reduced UDS or UV-sensitization by caffeine, whilst others showed marked sensitivity to EMS or MMC, and they may have mutations in different genes involved in nucleotide excision repair, post-replicational repair, base excision repair or recombinational repair. Previous mutants isolated as part of the FAECB collection have proved to be extremely important in characterizing mammalian DNA repair processes and cloning human repair genes and these current mutants, whilst not as hypersensitive to UV, may still have the potential to make further contributions.

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