Department of Biomedical Sciences and Pathobiology, VA-MD College of Veterinary Medicine, Virginia Tech, Blacksburg, Virginia, USA.
Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York, USA.
J Bacteriol. 2018 May 24;200(12). doi: 10.1128/JB.00105-18. Print 2018 Jun 15.
The YbeY endoribonuclease is one of the best-conserved proteins across the kingdoms of life. In the present study, we demonstrated that YbeY in is linked to a variety of important activities, including proper cellular morphology, mRNA transcript levels, and virulence. Deletion of in led to a small-colony phenotype when the bacteria were grown on agar medium, as well as to significant aberrations in the morphology of the bacterial cell as evidenced by electron microscopy. Additionally, compared to the parental strain, the Δ strain was significantly attenuated in both macrophage and mouse models of infection. The Δ strain also showed increased sensitivities to several -applied stressors, including bile acid, hydrogen peroxide, SDS, and paraquat. Transcriptomic analysis revealed that a multitude of mRNA transcripts are dysregulated in the Δ strain, and many of the identified mRNAs encode proteins involved in metabolism, nutrient transport, transcriptional regulation, and flagellum synthesis. We subsequently constructed gene deletion strains of the most highly dysregulated systems, and several of the YbeY-linked gene deletion strains exhibited defects in the ability of the bacteria to survive and replicate in primary murine macrophages. Taken together, these data establish a clear role for YbeY in the biology and virulence of ; moreover, this work further illuminates the highly varied roles of this widely conserved endoribonuclease in bacteria. spp. are highly efficient bacterial pathogens of animals and humans, causing significant morbidity and economic loss worldwide, and relapse of disease often occurs following antibiotic treatment of human brucellosis. As such, novel therapeutic strategies to combat infections are needed. Ribonucleases in the brucellae are understudied, and these enzymes represent elements that may be potential targets for future treatment approaches. The present work demonstrates the importance of the YbeY endoribonuclease for cellular morphology, efficient control of mRNA levels, and virulence in Overall, the results of this study advance our understanding of the critical roles of YbeY in the pathogenesis of the intracellular brucellae and expand our understanding of this highly conserved RNase.
YbeY 内切核糖核酸酶是生命王国中最保守的蛋白质之一。在本研究中,我们证明了 中的 YbeY 与多种重要的活性相关联,包括细胞形态、mRNA 转录水平和毒力。当细菌在琼脂培养基上生长时,缺失 中的 会导致小菌落表型,并且电镜观察表明细菌细胞形态存在显著异常。此外,与亲本菌株相比,Δ 菌株在巨噬细胞和小鼠感染模型中均显著减弱。Δ 菌株对几种施加的应激源,包括胆酸、过氧化氢、SDS 和百草枯,的敏感性也增加了。转录组分析显示,Δ 菌株中多种 mRNA 转录物失调,许多鉴定出的 mRNAs 编码参与代谢、营养物质运输、转录调节和鞭毛合成的蛋白质。我们随后构建了最高度失调系统的基因缺失菌株,并且 YbeY 相关基因缺失菌株中的几个在细菌在原代小鼠巨噬细胞中存活和复制的能力方面表现出缺陷。总之,这些数据确立了 YbeY 在 生物学和毒力中的明确作用;此外,这项工作进一步阐明了这种广泛保守的内切核糖核酸酶在细菌中的高度多样化作用。 spp. 是动物和人类的高效细菌病原体,在全球范围内造成严重的发病率和经济损失,并且在人类布鲁氏菌病用抗生素治疗后经常复发。因此,需要新的治疗策略来对抗 感染。布鲁氏菌中的核糖核酸酶研究不足,这些酶代表可能成为未来治疗方法的潜在靶点。本工作证明了 YbeY 内切核糖核酸酶在 细胞形态、有效控制 mRNA 水平和毒力中的重要性。总体而言,本研究结果提高了我们对 YbeY 在布鲁氏菌属细胞内发病机制中的关键作用的认识,并扩展了我们对这种高度保守的核糖核酸酶的理解。