Insertional Inactivation of Prevotella intermedia OxyR Results in Reduced Survival with Oxidative Stress and in the Presence of Host Cells.
Insertional Inactivation of Prevotella intermedia OxyR Results in Reduced Survival with Oxidative Stress and in the Presence of Host Cells.
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DOI：
10.3390/microorganisms9030551
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发表时间：
2021-03-07
期刊：
影响因子：
4.5
通讯作者：
Lewis JP
中科院分区：
文献类型：
--
作者：
Naito M;Belvin BR;Shoji M;Gui Q;Lewis JP
中间普氏菌是牙周机械治疗后出血患者龈下袋中最丰富的细菌之一。然而，尽管其丰富，其对牙周病的贡献的分子机制并不为人所知。这主要是由于缺乏遗传工具，可以检查预测的毒力因子在该细菌发病机制中的作用。在这里，我们报告的第一个突变体中的P. intermedia OMA 14菌株。该突变是编码推定OxyR调节剂的序列与编码厌氧细菌中大环内酯-林可酰胺抗性的ermF序列的等位基因交换替换。该突变体与真核细胞一起生长的能力严重受损，这表明它是干预策略的重要靶点。进一步的分析表明，它的生长能力与氧化应激物种，在过氧化氢和氧气的形式，受到严重影响。转录组分析表明，主要的失调基因编码的烷基氢过氧化物还原酶系统，AhpCF，介导保护过氧化氢胁迫。此外，编码Dps、CydA和Ftn的基因在突变株中下调，如使用qRT-PCR分析进一步验证的。总之，我们成功地产生了第一个中间青霉突变体，并表明OxyR缺陷型菌株不能与各种宿主细胞以及氧化应激一起存活。
One of the most abundant bacteria in the subgingival pockets of patients with bleeding following mechanical periodontal therapy is Prevotella intermedia. However, despite its abundance, the molecular mechanisms of its contribution to periodontal disease are not well known. This is mainly due to the lack of genetic tools that would allow examination of the role of predicted virulence factors in the pathogenesis of this bacterium. Here, we report on the first mutant in the P. intermedia OMA14 strain. The mutation is an allelic exchange replacement of the sequences coding for a putative OxyR regulator with ermF sequences coding for the macrolide–lincosamide resistance in anaerobic bacteria. The mutant is severely impaired in its ability to grow with eukaryotic cells, indicating that it is an important target for interventional strategies. Further analyses reveal that its ability to grow with oxidative stress species, in the form of hydrogen peroxide and oxygen, is severely affected. Transcriptome analysis reveals that the major deregulated genes code for the alkylhydroperoxide reductase system, AhpCF, mediating protection from peroxide stress. Moreover, genes coding for Dps, CydA and Ftn are downregulated in the mutant strain, as further verified using qRT-PCR analysis. In conclusion, we succeeded in generating the first P. intermedia mutant and show that the OxyR-deficient strain is unable to survive with a variety of host cells as well as with oxidative stress.
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DOI：
10.1093/dnares/dsv032
发表时间：
2016-02
期刊：
DNA research : an international journal for rapid publication of reports on genes and genomes
影响因子：
--
作者：
Naito M;Ogura Y;Itoh T;Shoji M;Okamoto M;Hayashi T;Nakayama K
通讯作者：
Nakayama K
DOI：
10.1073/pnas.1701587114
发表时间：
2017-08-15
影响因子：
11.1
作者：
Khademian, Maryam;Imlay, James A.
通讯作者：
Imlay, James A.
影响因子：
4.2
作者：
Baumgartner, JC;Watkins, BJ;Xia, T
通讯作者：
Xia, T
影响因子：
2.8
作者：
Olczak, Teresa;Sroka, Aneta;Olczak, Mariusz
通讯作者：
Olczak, Mariusz
影响因子：
2.6
作者：
Maeda, N;Okamoto, M;Fujita, H
通讯作者：
Fujita, H