DNA Phosphorothioate Modification Plays a Role in Peroxides Resistance in Streptomyces lividans.
DNA Phosphorothioate Modification Plays a Role in Peroxides Resistance in Streptomyces lividans.
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DNA 硫代磷酸酯修饰在浅青紫链霉菌的过氧化物抗性中发挥作用
DOI：
10.3389/fmicb.2016.01380
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发表时间：
2016
影响因子：
5.2
通讯作者：
Wang Z
中科院分区：
文献类型：
--
作者：
Dai D;Du A;Xiong K;Pu T;Zhou X;Deng Z;Liang J;He X;Wang Z
由dnd基因赋予的DNA磷硫酰化最初在土壤细菌变铅青链霉菌（Streptomyces lividans）中被发现，此后发现存在于多种细菌属中。然而，这种DNA骨架的硫修饰在变铅青链霉菌中的生理意义仍然未知。我们的研究表明，DNA磷硫酰化在该菌株对氧化应激的抗性中起主要作用。尽管链霉菌属表达多种过氧化氢酶/过氧化物酶和有机氢过氧化物抗性基因以保护自身免受过氧化物损伤，但在过氧化物处理后，变铅青链霉菌的野生型菌株与dnd⁻突变体相比，存活率高出2到10倍。RNA - seq实验表明，野生型菌株中过氧化氢酶和有机氢过氧化物抗性基因的表达没有上调，这表明对氧化应激的抗性不是由于DNA磷硫酰化对这些基因的上调作用。进行了定量RT - PCR分析以追踪过氧化物处理后过氧化氢酶和有机氢过氧化物抗性基因的表达。在对过氧化物的反应中，一系列这些基因在dnd⁻突变体中被激活，而不是在野生型菌株中。此外，在体内和体外，有机氢过氧化物过氧乙酸在有磷硫酰化修饰的情况下比没有该修饰时被更快地清除。dnd基因簇可被二硫键应激物二酰胺上调。总体而言，我们的观察结果表明，DNA磷硫酰化修饰在变铅青链霉菌中作为一种过氧化物抗性系统发挥作用。
DNA phosphorothioation, conferred by dnd genes, was originally discovered in the soil-dwelling bacterium Streptomyces lividans, and thereafter found to exist in various bacterial genera. However, the physiological significance of this sulfur modification of the DNA backbone remains unknown in S. lividans. Our studies indicate that DNA phosphorothioation has a major role in resistance to oxidative stress in the strain. Although Streptomyces species express multiple catalase/peroxidase and organic hydroperoxide resistance genes to protect them against peroxide damage, a wild type strain of S. lividans exhibited two-fold to 10-fold higher survival, compared to a dnd− mutant, following treatment with peroxides. RNA-seq experiments revealed that, catalase and organic hydroperoxide resistance gene expression were not up-regulated in the wild type strain, suggesting that the resistance to oxidative stress was not due to the up-regulation of these genes by DNA phosphorothioation. Quantitative RT-PCR analysis was conducted to trace the expression of the catalase and the organic hydroperoxide resistance genes after peroxides treatments. A bunch of these genes were activated in the dnd− mutant rather than the wild type strain in response to peroxides. Moreover, the organic hydroperoxide peracetic acid was scavenged more rapidly in the presence than in the absence of phosphorothioate modification, both in vivo and in vitro. The dnd gene cluster can be up-regulated by the disulfide stressor diamide. Overall, our observations suggest that DNA phosphorothioate modification functions as a peroxide resistance system in S. lividans.
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