DNA Strand Breakage by 125I-Decay in a Synthetic Oligodeoxynucleotide: Quantitative analysis of fragment distribution
The DNA breakage produced by decay of 125I in a double-stranded 41 bp oligodeoxynucleotide was investigated by DNA sequencing gel analysis. Use of both 5'- and 3'-end 32P labelling of the 125I containing strand provided the single-stranded breakage pattern at either side of the 125I-dC. Th...
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Zusammenfassung: | The DNA breakage produced by decay of 125I in a double-stranded 41 bp oligodeoxynucleotide was investigated by DNA sequencing gel analysis. Use of both 5'- and 3'-end 32P labelling of the 125I containing strand provided the single-stranded breakage pattern at either side of the 125I-dC. The asymmetric pattern of breakage relative to the 125I-labelled nucleotide enabled deconvolution of two components of breakage. One of them declines very quickly with nucleotide number from 125I-dC and dominates within 4-5 nucleotides. We assume this component to be associated with neutralisation of charged Te atom resulted from decay, or/and with radiation damage to an initial target other than the deoxyribosyl moiety-probably the bases. The second component depends on the geometrical distance between the 125I atom and deoxyribosyl atoms. These two components are responsible for breakage under conditions limiting radical mediated damage, namely in the presence of 2M dimethylsulphoxide. The third component, associated with radical-mediated damage, contributes to the total breakage during incubation in 20mM phosphate buffer alone, and under these incubation conditions dominates the breakage beyond 8-9 nucleotides from 125I-dC. The estimated average numbers of single-stranded breaks produced in the 125I-containing strand by each mechanism in 41-mer oligodeoxynucleotide with 125I-dC at 21st position are respectively 2.33, 0.98 and 0.63. |
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ISSN: | 0284-186X 1651-226X |
DOI: | 10.3109/02841869609104031 |