Deering R A, Jensen D S
Biophys J. 1973 Aug;13(8):780-94. doi: 10.1016/S0006-3495(73)86024-2.
Uptake of [methyl-(3)H]thymidine label from Escherichia coli 15T(-) into the DNA of Dictyostelium discoideum has been measured in control and [(60)Co]-gamma-irradiated cells of the resistant strain NC-4 (D(10), colony-forming survival = 300 krad) and two sensitive daughter strains, gammas-18 (D(10) = 75 krad) and gammas-13 (D(10) = 4 krad). Nuclear (n) and mitochondrial (m) DNA were resolved by isopycnic CsCl gradients. The uptake of label into n-DNA during the immediate postirradiation period was selectively inhibited by irradiation, compared with uptake into m-DNA. For all three strains, the gamma ray dose to reduce the uptake into n-DNA to 37% of the control during the first hour after irradiation was 3 krad, whereas for uptake into m-DNA it was 75 krad. After the initial dose- and strain-dependent lag, uptake into n-DNA resumed. gammas-18 showed longer lags in n-DNA synthesis and cell division than did NC-4. gammas-13 resumed n-DNA synthesis and cell division after slightly shorter lags than for NC-4. The early postlag uptake into n-DNA in this strain was almost at the control rate and was accompanied by division until the cell number had nearly doubled. The rate of label uptake then declined, division stopped, and gradual cell lysis ensued. The postdelay response of gammas-13 was almost independent of dose in the range of 10-100 krad. The response of gammas-18 in these and earlier experiments is consistent with the view-point that it is sensitive because of a decreased rate of repair of DNA damage. However, the basis for the sensitivity of gammas-13 seems to be more complex. This strain undergoes a premature but short-lived burst of n-DNA synthesis and division for what appears to be about one round of replication. Replication then ceases, even at very low doses, leading to greatly reduced probability of survival.
已在抗性菌株NC-4(D(10),集落形成存活率 = 300 krad)以及两个敏感子代菌株γs-18(D(10) = 75 krad)和γs-13(D(10) = 4 krad)的对照细胞和[(60)Co]-γ辐照细胞中,测量了[甲基-(3)H]胸苷标记从大肠杆菌15T(-)掺入盘基网柄菌DNA的情况。通过等密度CsCl梯度分离核(n)DNA和线粒体(m)DNA。与掺入m-DNA相比,辐照后即刻期间标记掺入n-DNA的过程受到辐照的选择性抑制。对于所有三个菌株,辐照后第一小时将n-DNA掺入量降低至对照的37%所需的γ射线剂量为3 krad,而对于掺入m-DNA则为75 krad。在最初的剂量和菌株依赖性延迟之后,n-DNA的掺入恢复。γs-18在n-DNA合成和细胞分裂方面的延迟比NC-4更长。γs-13在比NC-4稍短的延迟后恢复n-DNA合成和细胞分裂。该菌株延迟后早期n-DNA的掺入几乎以对照速率进行,并伴随着细胞分裂,直到细胞数量几乎翻倍。然后标记掺入速率下降,细胞分裂停止,随后细胞逐渐裂解。γs-13延迟后的反应在10 - 100 krad剂量范围内几乎与剂量无关。γs-18在这些及早期实验中的反应与以下观点一致,即它敏感是因为DNA损伤修复速率降低。然而,γs-13敏感的基础似乎更为复杂。该菌株经历了一次过早但短暂的n-DNA合成和分裂爆发,似乎进行了一轮复制。然后复制停止,即使在非常低的剂量下也是如此,导致存活概率大大降低。