Role of the Filifactor alocis Hypothetical Protein FA519 in Oxidative Stress Resistance.
Role of the Filifactor alocis Hypothetical Protein FA519 in Oxidative Stress Resistance.
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DOI：
10.1128/spectrum.01212-21
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发表时间：
2021-12-22
影响因子：
3.7
通讯作者：
Fletcher HM
中科院分区：
文献类型：
--
作者：
Aja E;Mishra A;Dou Y;Fletcher HM
在牙周袋中，环境条件、动态口腔微生物植物群和疾病之间存在直接相关性。口腔微环境中几种新发现的微生物物种的相对丰度提出了它们对疾病发展的影响的问题。一种这样的生物体，Filifactoralocis，对病原生物膜结构具有重要意义。此外，它的致病特性突出了其在牙周袋的氧化应激微环境中生存并改变微生物群落动态的能力。目前对其抗氧化应激机制及对致病性影响的认识还存在空白。在F. alocis在与牙龈卟啉单胞菌W83共同感染上皮细胞期间。生物信息学分析表明，FA 519含有一个Cys-X-X-Cys锌带结构域，该结构域可能与DNA结合和抗氧化应激有关。我们对FA 519进行了表征，以阐明其在F. alocis。与野生型菌株相比，F. Alocis等基因缺失突变体FLL 1013（Δ FA 519：：ermF）对过氧化氢和一氧化氮诱导的胁迫的敏感性显著降低。在同基因突变体中，形成生物膜和粘附并侵袭牙龈上皮细胞的能力也降低。重组FA 519蛋白具有还原过氧化氢和二硫键的内在能力，可保护DNA免受Fenton介导的损伤。总的来说，这些结果表明，FA 519参与氧化应激抗性，并可以调节重要的毒力属性在F。alocis。重要性Filifactoralocis是牙周社区的一个新兴成员，现在被提议作为牙周病的诊断指标。然而，由于缺乏可用于研究这种生物的遗传工具，对其毒力属性知之甚少。探讨了F. Alocis是未知的。因此，识别F. alocis在牙周袋的氧化应激环境中生存将导致对其毒力调节的理解，这将有助于开发新的治疗方法来对抗牙周病的影响。本研究的重点是FA 519，一个假设的蛋白质在F。alocis是一种多功能蛋白，在活性氧解毒途径中起重要作用。总的来说，我们的研究结果表明，FA 519参与氧化应激抗性，并可以调节重要的毒力属性在F。alocis。
In the periodontal pocket, there is a direct correlation between environmental conditions, the dynamic oral microbial flora, and disease. The relative abundance of several newly recognized microbial species in the oral microenvironment has raised questions on their impact on disease development. One such organism, Filifactor alocis, is significant to the pathogenic biofilm structure. Moreover, its pathogenic characteristics are highlighted by its ability to survive in the oxidative-stress microenvironment of the periodontal pocket and alter the microbial community dynamics. There is a gap in our understanding of its mechanism(s) of oxidative stress resistance and impact on pathogenicity. Several proteins, including HMPRFF0389-00519 (FA519), were observed in high abundance in F. alocis during coinfection of epithelial cells with Porphyromonas gingivalis W83. Bioinformatics analysis shows that FA519 contains a “Cys-X-X-Cys zinc ribbon domain” which could be involved in DNA binding and oxidative stress resistance. We have characterized FA519 to elucidate its roles in the oxidative stress resistance and virulence of F. alocis. Compared to the wild-type strain, the F. alocis isogenic gene deletion mutant, FLL1013 (ΔFA519::ermF), showed significantly reduced sensitivity to hydrogen peroxide and nitric oxide-induced stress. The ability to form biofilm and adhere to and invade gingival epithelial cells was also reduced in the isogenic mutant. The recombinant FA519 protein was shown to protect DNA from Fenton-mediated damage with an intrinsic ability to reduce hydrogen peroxide and disulfide bonds. Collectively, these results suggest that FA519 is involved in oxidative stress resistance and can modulate important virulence attributes in F. alocis. IMPORTANCE Filifactor alocis is an emerging member of the periodontal community and is now proposed to be a diagnostic indicator of periodontal disease. However, due to the lack of genetic tools available to study this organism, not much is known about its virulence attributes. The mechanism(s) of oxidative stress resistance in F. alocis is unknown. Therefore, identifying the adaptive mechanisms utilized by F. alocis to survive in the oxidative stress environment of the periodontal pocket would lead to understanding its virulence regulation, which could help develop novel therapeutic treatments to combat the effects of periodontal disease. This study is focused on the characterization of FA519, a hypothetical protein in F. alocis, as a multifunctional protein that plays an important role in the reactive oxygen species-detoxification pathway. Collectively, our results suggest that FA519 is involved in oxidative stress resistance and can modulate important virulence attributes in F. alocis.