Lobachevsky P N, Martin R F
Molecular Radiation Biology Laboratory, Peter MacCallum Cancer Institute, Melbourne, Australia.
Acta Oncol. 1996;35(7):809-15. doi: 10.3109/02841869609104031.
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 20 mM 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.
通过DNA测序凝胶分析,研究了双链41bp寡脱氧核苷酸中125I衰变产生的DNA断裂情况。对含125I链的5'端和3'端进行32P标记,可得到125I-dC两侧的单链断裂模式。相对于125I标记核苷酸的不对称断裂模式,使得能够对两种断裂成分进行反卷积分析。其中一种成分随着与125I-dC的核苷酸数增加而迅速下降,在4-5个核苷酸范围内占主导。我们认为该成分与衰变产生的带电荷碲原子的中和有关,和/或与脱氧核糖部分以外的初始靶点(可能是碱基)的辐射损伤有关。第二种成分取决于125I原子与脱氧核糖原子之间的几何距离。在限制自由基介导损伤的条件下,即在存在2M二甲基亚砜的情况下,这两种成分导致断裂。第三种成分与自由基介导的损伤有关,在仅含20mM磷酸盐缓冲液的孵育过程中对总断裂有贡献,在这些孵育条件下,在距125I-dC超过8-9个核苷酸处的断裂中占主导。在第21位含有125I-dC的41聚体寡核苷酸中,每种机制在含125I链中产生的单链断裂的估计平均数分别为2.33、0.98和0.63。