Department of Life Sciences, Albstadt-Sigmaringen University of Applied Sciences, Anton-Günther-Str. 51, 72488, Sigmaringen, Germany.
Department of Life Sciences, Albstadt-Sigmaringen University of Applied Sciences, Anton-Günther-Str. 51, 72488, Sigmaringen, Germany.
DNA Repair (Amst). 2020 Mar;87:102784. doi: 10.1016/j.dnarep.2019.102784. Epub 2020 Jan 3.
The DNA of human cells suffers about 1.000-100.000 oxidative lesions per day. One of the most common defects in this category is represented by 7,8-dihydro-8-oxoguanine. There are numerous exogenous effects on DNA that induce the intracellular generation of 7, 8-dihydro-8-oxoguanine. Therefore, a quantitatively sufficient repair of all occurring oxidative damaged guanine bases is often only partially feasible, especially in advanced age. Inadequate removal of these damages can subsequently lead to mutations and thus to serious diseases. All these aspects represent a dangerous situation for an organism. However, it is suspected that the amount of the 8-oxoguanine DNA glycosylase can be actively regulated on the level of gene expression by the redox-active properties of ubiquinol and thus its protein expression can be controlled. Using an real-time base excision repair assay including a melting curve analysis, the activity of the human 8-oxoguanine DNA glycosylase 1 was measured under the influence of ubiquinol. It was possible to observe a concentration-dependent increase in the activity of the 8-oxoguanine DNA glycosylase 1 under the influence of ubiquinol for the first time, both on purified and commercially acquired enzyme as well as on enzyme isolated from mitochondria of human fibroblasts. An increase in activity of this enzyme based on a change in cellular redox state caused by ubiquinol could not be confirmed. In addition, an increased gene expression of 8-oxoguanine-DNA glycosylase 1 under ubiquinol could not be observed. However, there was a change in bifunctionality in favor of an increased N-glycosylase activity and a direct interaction between ubiquinol and 8-oxoguanine DNA glycosylase 1. We suggest that ubiquinol contributes to the dissolution of a human 8-oxoguanine DNA glycosylase 1 end-product complex that forms after cutting into the sugar-phosphate backbone of the DNA with the resulting unsaturated 3'-phospho-α, β-aldehyde end and thereby inhibits further enzymatic steps.
人类细胞的 DNA 每天会遭受约 1000-100000 种氧化损伤。在这一类中最常见的缺陷之一是 7,8-二氢-8-氧鸟嘌呤。有许多外源性因素会导致细胞内产生 7,8-二氢-8-氧鸟嘌呤,从而对 DNA 造成影响。因此,定量地修复所有发生的氧化损伤鸟嘌呤碱基通常是部分可行的,尤其是在高龄时。这些损伤的清除不足随后可能导致突变,从而导致严重疾病。所有这些方面都对生物体构成了危险。然而,人们怀疑 8-氧鸟嘌呤 DNA 糖基化酶的量可以通过泛醌的氧化还原活性在基因表达水平上被主动调节,从而控制其蛋白质表达。通过使用包括熔解曲线分析的实时碱基切除修复测定法,在泛醌的影响下测量了人 8-氧鸟嘌呤 DNA 糖基化酶 1 的活性。首次观察到,在纯化酶和商业获得的酶以及从人成纤维细胞线粒体分离的酶中,在泛醌的影响下,8-氧鸟嘌呤 DNA 糖基化酶 1 的活性呈浓度依赖性增加。不能证实基于泛醌引起的细胞氧化还原状态变化的这种酶的活性增加。此外,不能观察到 8-氧鸟嘌呤-DNA 糖基化酶 1 的基因表达增加。然而,观察到双功能性的变化有利于增加 N-糖基化酶活性,并且泛醌和 8-氧鸟嘌呤 DNA 糖基化酶 1 之间存在直接相互作用。我们建议,泛醌有助于溶解人 8-氧鸟嘌呤 DNA 糖基化酶 1 末端产物复合物,该复合物在切入 DNA 的糖磷酸骨架后形成,其结果是不饱和 3'-磷酸-α,β-醛末端,并由此抑制进一步的酶促步骤。