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Abundance of the δ Subunit of RNA Polymerase Is Linked to the Virulence of Streptococcus agalactiae

机译:RNA聚合酶δ亚基的丰度与无乳链球菌的毒力有关。

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摘要

Group B streptococcus (GBS) remains a major cause of morbidity and mortality among newborn children. The bacterium is a commensal organism colonizing the rectum and the gastrointestinal and urogenital tracts of adults, but it can be transmitted to neonates by an ascending infection of the maternal genital tract or during parturition. We previously reported that a transposon insertion disrupting rpoE resulted in the decreased survival of the mutant in the neonatal rat sepsis model of GBS infection. rpoE encodes the δ protein, a subunit of RNA polymerase (RNAP) that has been characterized in Bacillus species. In this study, we confirm the association of the δ protein with purified GBS RNAP and show that it is expressed in strains representing all nine serotypes. Flow cytometric analysis of a reporter strain containing a transcriptional fusion of the rpoE promoter to gfp revealed that, in vitro, this gene is continuously expressed. Analysis of δ expression in the transposon mutant by quantitative Western blotting revealed a 10-fold reduction in relative abundance (which was linked to the attenuation in virulence that was observed for this mutant) compared to that for the wild-type strain. These data suggest that a minimum intracellular concentration of δ is necessary for this organism to cause disease.
机译:B组链球菌(GBS)仍然是新生婴儿发病和死亡的主要原因。该细菌是在成年人的直肠以及胃肠道和泌尿生殖道上定殖的一种共生生物,但是它可以通过孕妇生殖道的不断上升的感染或分娩而传播给新生儿。我们先前曾报道转座子插入破坏rpoE导致该突变体在GBS感染的新生大鼠败血症模型中的存活率降低。 rpoE编码δ蛋白,这是一种已在芽孢杆菌属中表征的RNA聚合酶(RNAP)的亚基。在这项研究中,我们证实了δ蛋白与纯化的GBS RNAP的关联,并表明它在代表所有9种血清型的菌株中表达。含有rpoE启动子与gfp转录融合的报告基因菌株的流式细胞分析表明,该基因在体外连续表达。通过定量蛋白质印迹分析转座子突变体中的δ表达,发现与野生型菌株相比,相对丰度降低了10倍(这与该突变体观察到的毒力降低有关)。这些数据表明,该生物体引起疾病所必需的最小细胞内δ浓度。

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