Molecular markers of ionizing radiation-induced gene mutations in mammalian cells

Abraham W. Hsie, Ronald C. Porter, Zhidong Xu, Yongjia Yu, Juan Sun, Martin L. Meltz, Jeffrey L. Schwartz

Research output: Contribution to journalArticle

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Abstract

We have isolated independent Chinese hamster ovary (CHO) cell mutants at the hypoxanthine guanine phosphoribosyltransferase (hprt) locus from untreated, 60Co γ-ray-exposed, and 212Bi α-exposed cells and identified the molecular changes underlying the mutation determined by multiplex polymerase chain reaction (PCR)-based exon deletion analysis. Both the parental CHO-K1 cells and the X-ray-sensitive mutant xrs-5 cells were studied. The radiosensitive xrs-5 cells are defective in DNA double-strand break rejoining ability and in V(D)J recombination, which can be complemented by Ku protein. Of the 71 spontaneous CHO-K1 hprt mutants analyzed, 78% showed no change in exon number or size, 20% showed loss of one to eight exons (partial deletion), and 3% showed loss of all nine hprt exons (total deletion). Exposure of CHO-K1 cells to 6 Gy of γ rays, which reduced survival levels to 10%, produced a high deletion spectrum with 45% of the 20 mutants analyzed showing a loss of one to eight exons and 30% showing total deletion. Exposure to an equitoxic dose of a radiation from 212Bi, a 220Rn daughter, resulted in a spectrum similar to the γ-ray spectrum in that 75% of the 49 mutants analyzed were deletions. The ex radiation, however, tended to produce larger intragenic deletions than γ radiation. Of the 92 spontaneous xrs-5 mutants analyzed for deletions, 43% showed a loss of one to eight exons and 14% showed total deletion. This suggests that, in certain regions of the hprt gene, base alterations can be converted into large deletions and alteration in the Ku protein complex can influence this type of mutational process. Exposure to α radiation (10% survival) to xrs-5 cells resulted in a deletion spectrum similar to that seen in CHO-K1 cells. Of the 49 mutants analyzed, 43% showed no change in exon number or size, 16% showed a loss of one to eight exons, and 41% showed total deletion. While the defect in xrs-5 cells has a profound effect on spontaneous mutant spectra, this defect does not appear to affect α-induced mutation spectra.

Original languageEnglish (US)
Pages (from-to)675-678
Number of pages4
JournalEnvironmental Health Perspectives
Volume104
Issue numberSUPPL. 3
StatePublished - May 1996

Fingerprint

Ionizing radiation
Ionizing Radiation
mutation
Exons
Genes
Cells
Hypoxanthine Phosphoribosyltransferase
Mutation
gene
Cricetulus
Ovary
Radiation
defect
V(D)J Recombination
Defects
protein
mutant
marker
ionising radiation
Polymerase chain reaction

Keywords

  • α radiation
  • γ radiation
  • DNA double-strand breaks
  • Gene mutation
  • Molecular mutagenesis
  • Radiation mutagenesis

ASJC Scopus subject areas

  • Environmental Science(all)
  • Environmental Chemistry
  • Public Health, Environmental and Occupational Health

Cite this

Hsie, A. W., Porter, R. C., Xu, Z., Yu, Y., Sun, J., Meltz, M. L., & Schwartz, J. L. (1996). Molecular markers of ionizing radiation-induced gene mutations in mammalian cells. Environmental Health Perspectives, 104(SUPPL. 3), 675-678.

Molecular markers of ionizing radiation-induced gene mutations in mammalian cells. / Hsie, Abraham W.; Porter, Ronald C.; Xu, Zhidong; Yu, Yongjia; Sun, Juan; Meltz, Martin L.; Schwartz, Jeffrey L.

In: Environmental Health Perspectives, Vol. 104, No. SUPPL. 3, 05.1996, p. 675-678.

Research output: Contribution to journalArticle

Hsie, AW, Porter, RC, Xu, Z, Yu, Y, Sun, J, Meltz, ML & Schwartz, JL 1996, 'Molecular markers of ionizing radiation-induced gene mutations in mammalian cells', Environmental Health Perspectives, vol. 104, no. SUPPL. 3, pp. 675-678.
Hsie AW, Porter RC, Xu Z, Yu Y, Sun J, Meltz ML et al. Molecular markers of ionizing radiation-induced gene mutations in mammalian cells. Environmental Health Perspectives. 1996 May;104(SUPPL. 3):675-678.
Hsie, Abraham W. ; Porter, Ronald C. ; Xu, Zhidong ; Yu, Yongjia ; Sun, Juan ; Meltz, Martin L. ; Schwartz, Jeffrey L. / Molecular markers of ionizing radiation-induced gene mutations in mammalian cells. In: Environmental Health Perspectives. 1996 ; Vol. 104, No. SUPPL. 3. pp. 675-678.
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AU - Schwartz, Jeffrey L.

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N2 - We have isolated independent Chinese hamster ovary (CHO) cell mutants at the hypoxanthine guanine phosphoribosyltransferase (hprt) locus from untreated, 60Co γ-ray-exposed, and 212Bi α-exposed cells and identified the molecular changes underlying the mutation determined by multiplex polymerase chain reaction (PCR)-based exon deletion analysis. Both the parental CHO-K1 cells and the X-ray-sensitive mutant xrs-5 cells were studied. The radiosensitive xrs-5 cells are defective in DNA double-strand break rejoining ability and in V(D)J recombination, which can be complemented by Ku protein. Of the 71 spontaneous CHO-K1 hprt mutants analyzed, 78% showed no change in exon number or size, 20% showed loss of one to eight exons (partial deletion), and 3% showed loss of all nine hprt exons (total deletion). Exposure of CHO-K1 cells to 6 Gy of γ rays, which reduced survival levels to 10%, produced a high deletion spectrum with 45% of the 20 mutants analyzed showing a loss of one to eight exons and 30% showing total deletion. Exposure to an equitoxic dose of a radiation from 212Bi, a 220Rn daughter, resulted in a spectrum similar to the γ-ray spectrum in that 75% of the 49 mutants analyzed were deletions. The ex radiation, however, tended to produce larger intragenic deletions than γ radiation. Of the 92 spontaneous xrs-5 mutants analyzed for deletions, 43% showed a loss of one to eight exons and 14% showed total deletion. This suggests that, in certain regions of the hprt gene, base alterations can be converted into large deletions and alteration in the Ku protein complex can influence this type of mutational process. Exposure to α radiation (10% survival) to xrs-5 cells resulted in a deletion spectrum similar to that seen in CHO-K1 cells. Of the 49 mutants analyzed, 43% showed no change in exon number or size, 16% showed a loss of one to eight exons, and 41% showed total deletion. While the defect in xrs-5 cells has a profound effect on spontaneous mutant spectra, this defect does not appear to affect α-induced mutation spectra.

AB - We have isolated independent Chinese hamster ovary (CHO) cell mutants at the hypoxanthine guanine phosphoribosyltransferase (hprt) locus from untreated, 60Co γ-ray-exposed, and 212Bi α-exposed cells and identified the molecular changes underlying the mutation determined by multiplex polymerase chain reaction (PCR)-based exon deletion analysis. Both the parental CHO-K1 cells and the X-ray-sensitive mutant xrs-5 cells were studied. The radiosensitive xrs-5 cells are defective in DNA double-strand break rejoining ability and in V(D)J recombination, which can be complemented by Ku protein. Of the 71 spontaneous CHO-K1 hprt mutants analyzed, 78% showed no change in exon number or size, 20% showed loss of one to eight exons (partial deletion), and 3% showed loss of all nine hprt exons (total deletion). Exposure of CHO-K1 cells to 6 Gy of γ rays, which reduced survival levels to 10%, produced a high deletion spectrum with 45% of the 20 mutants analyzed showing a loss of one to eight exons and 30% showing total deletion. Exposure to an equitoxic dose of a radiation from 212Bi, a 220Rn daughter, resulted in a spectrum similar to the γ-ray spectrum in that 75% of the 49 mutants analyzed were deletions. The ex radiation, however, tended to produce larger intragenic deletions than γ radiation. Of the 92 spontaneous xrs-5 mutants analyzed for deletions, 43% showed a loss of one to eight exons and 14% showed total deletion. This suggests that, in certain regions of the hprt gene, base alterations can be converted into large deletions and alteration in the Ku protein complex can influence this type of mutational process. Exposure to α radiation (10% survival) to xrs-5 cells resulted in a deletion spectrum similar to that seen in CHO-K1 cells. Of the 49 mutants analyzed, 43% showed no change in exon number or size, 16% showed a loss of one to eight exons, and 41% showed total deletion. While the defect in xrs-5 cells has a profound effect on spontaneous mutant spectra, this defect does not appear to affect α-induced mutation spectra.

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