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首页> 外文期刊>Biochemistry >Repression of Staphylococcus aureus SrrAB using inducible antisense srrA alters growth and virulence factor transcript levels
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Repression of Staphylococcus aureus SrrAB using inducible antisense srrA alters growth and virulence factor transcript levels

机译:使用诱导型反义srrA抑制金黄色葡萄球菌SrrAB改变生长和毒力因子转录水平

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摘要

Our laboratory has previously identified the staphylococcal respiratory response (SrrAB), a Staphylococcus aureus two-component system that acts in the global regulation of virulence factors. In strain RN4220, SrrAB downregulated production of agr RNAIII, protein A, and TSST-1, particularly under low-oxygen conditions. Work by another group showed that SrrAB regulates energy metabolism genes and indicated that SrrAB may regulate energy transduction in response to changes in oxygen availability. In this study we investigate the role of SrrAB in regulating RNAIII, tst, spa, icaR, and icaA in a clinical isolate of S. aureus, MN8. We employ an inducible antisense vector, pYJY4, in order to repress transcription of srrAB. Transcript levels were assessed by reverse transcription followed by quantitative PCR. Repression of srrAB in rich media under aerobic growth conditions shows that SrrAB is required for expression of tst, spa, and icaR transcripts at wild-type levels. Comparisons made between rich media under aerobic conditions vs low-oxygen conditions show that srrAB transcript levels are not altered by oxygen alone. Previous studies performed on strain RN4220 under low-oxygen conditions indicate that SrrAB represses tst and spa transcript when the amount of oxygen is limited. We propose that, under aerobic conditions, SrrAB enhances the levels of tst, spa, and icaR, while under low-oxygen conditions, SrrAB decreases the levels of these three transcripts.
机译:我们的实验室先前已经确定了葡萄球菌呼吸反应(SrrAB),这是一种金黄色葡萄球菌两组分系统,可在毒力因子的全球调控中发挥作用。在菌株RN4220中,SrrAB下调了agr RNAIII,蛋白A和TSST-1的产生,特别是在低氧条件下。另一个小组的工作表明,SrrAB调节能量代谢基因,并表明SrrAB可能响应氧可利用性的变化而调节能量转导。在这项研究中,我们调查了SrrAB在调节金黄色葡萄球菌MN8临床分离物中的RNAIII,tst,spa,icaR和icaA中的作用。为了抑制srrAB的转录,我们使用了诱导型反义载体pYJY4。通过逆转录然后定量PCR来评估转录物水平。在有氧生长条件下,在富培养基中抑制srrAB表明,在野生型水平上表达tst,spa和icaR转录本需要SrrAB。在有氧条件下和低氧条件下的富培养基之间进行的比较表明,单独的氧气不会改变srrAB转录水平。先前在低氧条件下对菌株RN4220进行的研究表明,当氧气量有限时,SrrAB会抑制tst和spa转录本。我们建议,在有氧条件下,SrrAB会提高tst,spa和icaR的水平,而在低氧条件下,SrrAB会降低这三个转录本的水平。

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