Protective role of the PG1036-PG1037-PG1038 operon in oxidative stress in Porphyromonas gingivalis W83.
Protective role of the PG1036-PG1037-PG1038 operon in oxidative stress in Porphyromonas gingivalis W83.
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DOI：
10.1371/journal.pone.0069645
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Fletcher HM
中科院分区：
文献类型：
--
作者：
Henry LG;Aruni W;Sandberg L;Fletcher HM
作为一种厌氧菌，牙龈卟啉单胞菌在初始定植和活动性牙周病期间明显受到牙周袋恶劣炎症环境的影响。我们之前报道过，牙龈卟啉单胞菌中涉及8-oxo-7,8-二氢鸟嘌呤（8-oxoG）的氧化应激诱导的DNA损伤的修复可能是通过一种未描述的机制发生的。 DNA 亲和分级分离鉴定出 PG1037（一种保守的假设蛋白），以及其他与 8-oxoG 损伤结合的蛋白。 PG1037 是牙龈卟啉单胞菌中 uvrA-PG1037-pcrA 操纵子的一部分，已知该操纵子在 H2O2 诱导的应激下上调。使用基于PCR的线性转化方法通过等位基因交换诱变使uvrA和pcrA基因失活。多次尝试灭活 PG1037 均未成功。与铺在布鲁氏菌血琼脂上的野生型相似，uvrA 和 pcrA 缺陷突变体呈黑色且呈 β 溶血性。与亲本菌株相比，这些同基因突变体的牙龈蛋白酶活性也降低，并且对 H2O2 和紫外线照射更敏感。此外，糖基化酶测定表明，野生型和突变型牙龈卟啉单胞菌菌株的 8-oxoG 修复活性相似。多种蛋白质（其中一些已知具有氧化还原酶活性）已被证明与 PG1037 相互作用。纯化的重组PG1037蛋白可以保护DNA免受H2O2引起的损伤。总的来说，这些发现表明 uvrA-PG1037-pcrA 操纵子可能在牙龈卟啉单胞菌过氧化氢应激诱导的抗性中发挥重要作用。
As an anaerobe, Porphyromonas gingivalis is significantly affected by the harsh inflammatory environment of the periodontal pocket during initial colonization and active periodontal disease. We reported previously that the repair of oxidative stress-induced DNA damage involving 8-oxo-7,8-dihydroguanine (8-oxoG) may occur by an undescribed mechanism in P. gingivalis. DNA affinity fractionation identified PG1037, a conserved hypothetical protein, among other proteins, that were bound to the 8-oxoG lesion. PG1037 is part of the uvrA-PG1037-pcrA operon in P. gingivalis which is known to be upregulated under H2O2 induced stress. A PCR-based linear transformation method was used to inactivate the uvrA and pcrA genes by allelic exchange mutagenesis. Several attempts to inactivate PG1037 were unsuccessful. Similar to the wild-type when plated on Brucella blood agar, the uvrA and pcrA-defective mutants were black-pigmented and beta-hemolytic. These isogenic mutants also had reduced gingipain activities and were more sensitive to H2O2 and UV irradiation compared to the parent strain. Additionally, glycosylase assays revealed that 8-oxoG repair activities were similar in both wild-type and mutant P. gingivalis strains. Several proteins, some of which are known to have oxidoreducatse activity, were shown to interact with PG1037. The purified recombinant PG1037 protein could protect DNA from H2O2-induced damage. Collectively, these findings suggest that the uvrA-PG1037-pcrA operon may play an important role in hydrogen peroxide stress-induced resistance in P. gingivalis.
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影响因子：
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