Frameshift mutation in PRKDC, the gene for DNA-PKcs, in the DNA repair-defective, human, glioma-derived cell line M059J
Frameshift mutation in PRKDC, the gene for DNA-PKcs, in the DNA repair-defective, human, glioma-derived cell line M059J
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DOI：
10.1667/0033-7587(2001)156
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发表时间：
2001-07-01
影响因子：
3.4
通讯作者：
Allalunis-Turner, MJ
中科院分区：
文献类型：
--
作者：
Anderson, CW;Dunn, JJ;Allalunis-Turner, MJ
胶质瘤细胞系M059J对电离辐射敏感，缺乏DNA-PK活性，不表达催化亚单位DNA-PKcs的蛋白，而来自同一肿瘤的姊妹细胞系M059K具有正常的DNA-PK活性。这两个细胞系都接近五倍体，并且都有8号染色体的多个副本，DNA-PKcs基因PRKDC位于8号染色体上。对扩增出的外显子进行序列分析，发现M059J细胞中缺失与PRKDC密码子1351(ACC，Thr)的第一个核苷酸相对应的外显子32的一个“A”核苷酸。M059K细胞的DNA只有野生型序列，缺失A核苷酸会在33号外显子早期终止DNA-PKcs阅读框。对87名无关个体的PRKDC外显子32周围序列的分析表明，除了该外显子3‘端附近的一个三联体重复外，没有发现其他核苷酸的多态；在外显子32上没有个体发生移码突变。在M059J和M059K细胞的PRKDC中，没有观察到类似于15000bp的基因组序列的其他序列差异，包括外显子5到38及其周围的内含子序列，这表明在获得导致M059J细胞系的突变之前，该基因座的纯合性可能降低。(C)2001年，由辐射研究学会提供。
The glioma-derived cell line M059J is hypersensitive to ionizing radiation, lacks DNA-PK activity, and fails to express protein for the catalytic subunit, DNA-PKcs, while a sister cell line, M059K, derived from the same tumor, has normal DNA-PK activity. Both cell lines are near pentaploid and have multiple copies of chromosome 8, the chromosome on which the DNA-PKcs gene, PRKDC, is located. Sequence analysis of PCR-amplified exons revealed the loss in M059J cells of a single "A" nucleotide in exon 32, corresponding to the first nucleotide of codon 1351 (ACC, Thr) of PRKDC. Loss of the "A" nucleotide would terminate the DNA-PKcs reading frame early in exon 33, DNA from M059K cells had only the wild-type sequence. An analysis of sequences surrounding PRKDC ex-on 32 from 87 unrelated individuals revealed no polymorphic nucleotides except for a triplet repeat near the 3' end of this exon; no individual had a frameshift mutation in exon 32. No other sequence differences in PRKDC between M059J and M059K cells were observed in similar to 15,000 bp of genomic sequence including the sequences of exons 5 through 38 and surrounding intron sequence, suggesting a possible reduction to homozygosity at this locus prior to acquisition of the mutation leading to the M059J cell line. (C) 2001 by Radiation Research Society.