Esipova V V, Lisovskaia K V, Kriviskiĭ A S
Genetika. 1978 Jun;14(6):975-86.
Mutagenesis in extracellular phage sd by 8-metoxypsoralen (8-MOP) and longwave (lambda greater than 310 nm) UV-irradiation has been established. The kinetics of lethal and mutagenic effects of 8-MOP+light was studied. The efficiency of mutagenesis on the first linear part of mutation curve was 0.3% per the lethal hit which is 2 times lower than that of shortwave (lambda=254 nm) UV-irradiation. The maximum yield of mutants makes up 1%, after which the mutation curve is maintained. It has been established that the main (may be the only) contribution into mutagenesis is made by monoadducts, whereas the lethal effect is conditioned by diadducts (cross-links). The comparison of the efficiency of mutagenic effects of 8-MOP+light with mutagenic effects of other kinds of irradiations was carried out. The possibility of repair of damaged 8-MOP+light phage sd DNA by transfection of Escherichia coli C (uvr+) and Cs (uvr-) lysozyme spheroplasts has been established. The repair mechanism of photodamage in intact phage sd induced with 8-MOP+light was also investigated using the method of two-step irradiation. It has been shown that 65% of photodamages are repaired in E. coli SK cells in the M9 medium, i. e. under cellular metabolism. The recovery of phage sd is completely inhibited in phosphate buffer. Unlike chloramphenicol (150 microgram/ml), 1% caffeine blocks the phage recovery only by 30%. The participation of phage sd determining enzymes in its intracellular recovery from 8-MOP+light damages is assumed.
已证实8-甲氧基补骨脂素(8-MOP)和长波(λ>310nm)紫外线照射可诱导细胞外噬菌体sd发生诱变。研究了8-MOP+光的致死和诱变效应动力学。在突变曲线的第一段线性部分,诱变效率为每致死一击0.3%,这比短波(λ=254nm)紫外线照射低2倍。突变体的最大产量为1%,此后突变曲线保持稳定。已确定单加合物对诱变起主要(可能是唯一)作用,而致死效应则由双加合物(交联)引起。对8-MOP+光的诱变效应与其他类型辐射的诱变效应进行了比较。通过转染大肠杆菌C(uvr+)和Cs(uvr-)溶菌酶原生质球,确定了受损的8-MOP+光噬菌体sd DNA的修复可能性。还采用两步照射法研究了完整噬菌体sd中8-MOP+光诱导的光损伤修复机制。结果表明,在M9培养基中,即在细胞代谢条件下,大肠杆菌SK细胞中65%的光损伤得到修复。在磷酸盐缓冲液中,噬菌体sd的恢复完全受到抑制。与氯霉素(150μg/ml)不同,1%的咖啡因仅使噬菌体恢复率降低30%。推测噬菌体sd决定酶参与其从8-MOP+光损伤中的细胞内恢复过程。