Griffiths T D, Ling S Y
Biochem Biophys Res Commun. 1984 Sep 17;123(2):742-8. doi: 10.1016/0006-291x(84)90292-4.
It has been reported that changes in deoxynucleotide pool levels following exposure to UV might lead to an overestimation of the UV-induced depression in DNA synthesis as analyzed by incorporation of 3H-thymidine (1). We attempted to determine the importance of such pool effects in two ways. First, we examined the grain density along DNA fiber autoradiographs obtained from CHO AA8, and CHO UV-5 cells exposed or sham exposed to UV. Exposure to UV did not alter the grain density immediately or 5 hours after exposure to UV in these cell lines. Second, we examined the kinetics of incorporation of 3H-dTTP in permeabilized AA8 cells under conditions of increased dCTP or increased exogenous dTTP levels. The extent of the depression of 3H-dTTP was identical under all incubation conditions.
据报道,暴露于紫外线后脱氧核苷酸库水平的变化可能导致通过掺入³H-胸腺嘧啶核苷分析的紫外线诱导的DNA合成抑制的高估(1)。我们试图通过两种方式确定这种库效应的重要性。首先,我们检查了从暴露或假暴露于紫外线的CHO AA8和CHO UV-5细胞获得的DNA纤维放射自显影片上的颗粒密度。在这些细胞系中,暴露于紫外线后立即或5小时后,颗粒密度没有改变。其次,我们在dCTP增加或外源性dTTP水平增加的条件下,检查了通透的AA8细胞中³H-dTTP掺入的动力学。在所有孵育条件下,³H-dTTP的抑制程度是相同的。