Frameshift mutation in PRKDC, the gene for DNA-PKcs, in the DNA repair-defective, human, glioma-derived cell line M059J

• Published in: Radiation research Vol. 156; no. 1; p. 2
• Main Authors: Anderson, C W, Dunn, J J, Freimuth, P I, Galloway, A M, Allalunis-Turner, M J
• Format: Journal Article
• Language: English
• Published: United States 01.07.2001
• Subjects: Catalytic Domain - genetics; Chromosomes, Human, Pair 8 - genetics; DNA Mutational Analysis; DNA Repair - genetics; DNA-Activated Protein Kinase; DNA-Binding Proteins; Exons - genetics; Frameshift Mutation - genetics; Glioma - enzymology; Humans; Male; Molecular Sequence Data; Nuclear Proteins; Polymerase Chain Reaction; Polyploidy; Protein Subunits; Protein-Serine-Threonine Kinases - genetics; Radiation Tolerance - genetics; Sequence Analysis, DNA; Tumor Cells, Cultured
• ISSN: 0033-7587
• DOI: 10.1667/0033-7587(2001)156[0002:FMIPTG]2.0.CO;2

Anderson, C. W., Dunn, J. J., Freimuth, P. I., Galloway, A. M. and Allalunis-Turner, M. J. Frameshift Mutation in PRKDC, the Gene for DNA-PKcs, in the DNA Repair-Defective, Human, Glioma-Derived Cell Line M059J. Radiat. Res. 156, 2-9 (2001). 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 exon 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 approximately 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.