Braun J E, Wanamarta A H, van den Akker E, Lafleur M V, Retèl J
Department of Medical Oncology, Vrije Universiteit, Amsterdam, The Netherlands.
Mutat Res. 1993 Oct;289(2):255-63. doi: 10.1016/0027-5107(93)90076-r.
To get more insight into the possible mutagenic consequences of DNA damage induced by radiation-generated H radicals (.H), a nitrogen-saturated solution of double-stranded (ds) M13mp10 DNA in phosphate buffer was irradiated with gamma-rays. Under these conditions 55% of the DNA-damaging species consists of H radicals and 45% of OH radicals (.OH). The mutations were investigated in a 144-bp mutational target sequence inserted into the lacZ alpha gene. A very specific mutation spectrum was obtained with respect to the type of mutations. Twenty out of the 28 radiation-induced mutations were C/G to A/T transversions; the remaining 8 mutations were 4 C/G to G/C transversions, 2 C/G to T/A transitions, one T/A to A/T transversion and only one -1 bp deletion. The mutations were rather randomly distributed along the 144-bp mutation target sequence with no clear mutational hot spots. When these results are compared with those we have obtained previously after irradiation of ds M13mp10 DNA under O2 (100% .OH) or N2O (90% .OH; 10% .H) (Hoebee et al., 1988, 1989), the data strongly suggest that H radicals may be responsible for the observed C/G to A/T transversions but not for -1 bp deletions.
为了更深入了解由辐射产生的氢自由基(·H)诱导的DNA损伤可能产生的诱变后果,用γ射线照射磷酸盐缓冲液中双链(ds)M13mp10 DNA的氮气饱和溶液。在这些条件下,55%的DNA损伤物种由氢自由基组成,45%由羟基自由基(·OH)组成。在插入lacZα基因的144 bp突变靶序列中研究突变。就突变类型而言,获得了非常特异的突变谱。28个辐射诱导的突变中有20个是C/G到A/T的颠换;其余8个突变是4个C/G到G/C的颠换、2个C/G到T/A的转换、1个T/A到A/T的颠换以及仅1个-1 bp的缺失。这些突变在144 bp的突变靶序列上相当随机地分布,没有明显的突变热点。当将这些结果与我们之前在O2(100%·OH)或N2O(90%·OH;10%·H)照射ds M13mp10 DNA后获得的结果进行比较时(Hoebee等人,1988年,1989年),数据强烈表明氢自由基可能是观察到的C/G到A/T颠换的原因,但不是-1 bp缺失的原因。