Kemp L M, Sedgwick S G, Jeggo P A
Mutat Res. 1984 Nov-Dec;132(5-6):189-96. doi: 10.1016/0167-8817(84)90037-3.
Six CHO mutants have previously been described as being sensitive to ionizing radiation and bleomycin treatment, with little or no cross sensitivity to UV-radiation (Jeggo and Kemp, 1983). Their ability to rejoin single- and double-strand breaks has been examined here. Using two techniques, gradient sedimentation and alkaline elution, no difference could be observed between wild-type and mutant strains in the initial number of single-strand breaks induced, the rate of rejoining, or the final level of single-strand breaks rejoined. Thus, a major inability to rejoin single-strand breaks is not the basis for sensitivity in these mutants. In contrast, all 6 mutants showed a decreased ability to rejoin the double-strand breaks induced by gamma-irradiation as measured by neutral elution. Rejoining of half of the breaks occurred in 37 min in wild-type cells and reached a maximum level of 72% after 2 h. All the mutants showed a decreased rate of rejoining, and the final level was 17% of that observed in the wild-type in the most defective mutant, and ranged from 35 to 69% in the other 5 mutants. These are the first mammalian cell mutants to be described with a defect in double-strand break rejoining.
之前已经描述过六个中国仓鼠卵巢细胞(CHO)突变体对电离辐射和博来霉素治疗敏感,对紫外线辐射几乎没有交叉敏感性(杰戈和肯普,1983年)。在此研究了它们重新连接单链和双链断裂的能力。使用梯度沉降和碱性洗脱这两种技术,在诱导的单链断裂的初始数量、重新连接的速率或重新连接的单链断裂的最终水平方面,未观察到野生型和突变株之间存在差异。因此,这些突变体中重新连接单链断裂的主要缺陷不是其敏感性的基础。相比之下,通过中性洗脱测量,所有6个突变体在重新连接由γ射线诱导的双链断裂方面能力下降。野生型细胞中一半的断裂在37分钟内重新连接,2小时后达到72%的最高水平。所有突变体的重新连接速率均下降,在缺陷最严重的突变体中,最终水平为野生型的17%,在其他5个突变体中,该水平在35%至69%之间。这些是首批被描述为双链断裂重新连接存在缺陷的哺乳动物细胞突变体。