Krisch R E, Flick M B
Department of Radiation Oncology, University of Pennsylvania School of Medicine, Philadelphia 19104.
Radiat Res. 1988 Dec;116(3):462-71.
We have previously published the techniques and preliminary results of an SV40 viral probe assay for gamma-radiation-induced single- and double-strand DNA breaks and their intracellular repair in higher cells (Radiat. Res. 101, 356-372, 1985). Those experiments with SV40 infected CV-1 monkey kidney cells suggested that this assay technique demonstrates slow but extensive intracellular repair of single-strand breaks (SSB), and possible early repair of double-strand breaks (DSB), followed by later induction of DSB. Following up on these early observations, many additional infection-incubation experiments have now been performed with both human and simian cells. Analysis of data from these experiments involving up to 6 h of postinfection intranuclear incubation shows the same distribution of strand break damage in incubated and unincubated samples. This implies that under these experimental conditions there is neither intracellular repair nor further production of SSB or DSB in intranuclear viral DNA. We have evidence which suggests that this lack of repair or degradation occurs because the bulk of intranuclear SV40 DNA is relatively inaccessible to host cell enzymes.
我们之前已经发表了一种用于检测γ辐射诱导的高等细胞中单链和双链DNA断裂及其细胞内修复的SV40病毒探针检测技术和初步结果(《辐射研究》101, 356 - 372, 1985)。那些用SV40感染CV - 1猴肾细胞的实验表明,这种检测技术显示出单链断裂(SSB)的细胞内修复缓慢但广泛,双链断裂(DSB)可能早期修复,随后是后期诱导DSB。基于这些早期观察结果,现在已经对人类和猿猴细胞进行了许多额外的感染 - 孵育实验。对这些实验数据的分析涉及感染后长达6小时的核内孵育,结果显示孵育和未孵育样品中的链断裂损伤分布相同。这意味着在这些实验条件下,核内病毒DNA中既没有细胞内修复,也没有SSB或DSB的进一步产生。我们有证据表明,这种缺乏修复或降解的情况发生是因为大部分核内SV40 DNA相对难以被宿主细胞酶作用。