Phenotypic heterogeneity within the first complementation group of UV-sensitive mutants of Chinese hamster cell lines

Małgorzata Z. Zdzienicka , G.P. van der Schans , A. Westerveld , A.A. van Zeeland , J.W.I.M. Simons
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引用次数: 43

Abstract

A DNA-repair mutant was characterized that has the extraordinary and interesting properties of extreme sensitivity to UV killing combined with a high level of nucleotide excision repair. The mutant V-H1 isolated from the V79 Chinese hamster cell line appeared very stable, with a reversion frequency of about 3.5 × 10−7. Genetic complementation analysis indicates that V-H1 belongs to the first complementation group of UV-sensitive Chinese hamster ovary (CHO) mutants described by Thompson et al. (1981). This correponds with data on cross-sensitivity and mutation induction after UV irradiation published by this group. Surprisingly, the mutant V-H1 shows only slightly reduced (to ∼ 70%) unscheduled DNA synthesis (UDS) after UV exposure, while the other two mutants of this complementation group are deficient in UDS after UV. In agreement with the high residual UDS, in V-H1 also the amount of repair replication in response to UV treatment is relatively high (∼ 50%). It has also been shown that the incision step of the nucleotide excision pathway takes place in V-H1 (with a lower rate than observed in wild-type cells), whereas another mutant (UV5) of the same complementation group is deficient in incision.

This heterogeneity within the first complementation group indicates that the repair gene of this complementation group may have more than one functionally domain or that the gene is not involved in the incision per se but is involved in e.g. preferential repair of active genes.

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中国仓鼠细胞系紫外线敏感突变体第一互补组的表型异质性
一个dna修复突变体的特征是具有对紫外线杀灭的极端敏感性以及高水平的核苷酸切除修复的非凡和有趣的特性。从中国仓鼠V79细胞系中分离得到的突变体V-H1表现出非常稳定的特性,其逆转频率约为3.5 × 10−7。遗传互补分析表明,V-H1属于Thompson等(1981)描述的中国仓鼠卵巢紫外线敏感突变体(CHO)的第一个互补类群。这与该小组发表的紫外线照射后交叉敏感性和突变诱导的数据相一致。令人惊讶的是,突变体V-H1在紫外线照射后仅显示出轻微减少(约70%)的非预定DNA合成(UDS),而该互补组的其他两个突变体在紫外线照射后缺乏UDS。与高残留UDS一致,在V-H1中,对紫外线处理的修复复制量也相对较高(约50%)。研究还表明,核苷酸切除途径的切口步骤发生在V-H1中(比在野生型细胞中观察到的速率低),而同一互补组的另一个突变体(UV5)缺乏切口。第一互补组内的这种异质性表明,该互补组的修复基因可能具有多个功能域,或者该基因本身不参与切口,但参与活性基因的优先修复。
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