Laboratorio de Microbiología Molecular, Facultad de Ciencias Biológicas, Universidad Andres Bello, Santiago, Chile.
Great Lakes Bioenergy Research Center and Department of Biomolecular Chemistry, University of Wisconsin-Madison, Madison, WI, USA.
Biochem Biophys Res Commun. 2014 Jul 18;450(1):641-5. doi: 10.1016/j.bbrc.2014.06.031. Epub 2014 Jun 14.
Typically, the expression of sRNAs is activated in response to environmental stimuli in order to regulate gene expression through post-transcriptional mechanisms. In the present work we show that the Salmonellatyphimurium paralogous sRNAs RyhB-1 and RyhB-2 are induced in response to the nitrosating agent S-nitrosoglutathione (GSNO). Inactivation of these sRNAs decreased S. typhimurium resistance to GSNO and increased the levels of nitrosylated proteins. These results prompted us to evaluate a possible role of these sRNAs in nitrosative stress resistance. RNA profiling was used as a screen to identify novel RyhB-1 and RyhB-2 regulated targets. A subset of genes was filtered based on their potential role in the response to nitrosative stress and their expression was analyzed by quantitative RT-PCR in wild type, single and double mutant strains (ΔryhB1, ΔryhB2 and ΔryhB1 ΔryhB2) treated with GSNO. In response to GSNO RyhB-1 and RyhB-2 negatively regulate the expression of the genes cyoABC (cytochrome bo oxidase), cydB (cytochrome bd oxidase), cybC (cytochrome b-562), and positively regulate the nirBCD operon (nitrite reductase system). Together, these results suggest that RyhB-1 and RyhB-2 finely tune the expression of genes coding for cytochrome oxidases and the nitrate reductase system, allowing the cell to cope with GSNO-induced stress.
通常情况下,sRNAs 的表达会被环境刺激激活,从而通过转录后机制来调节基因表达。在本工作中,我们发现 Salmonellatyphimurium 的平行 sRNAs RyhB-1 和 RyhB-2 会响应亚硝化剂 S-亚硝基谷胱甘肽(GSNO)而被诱导。这些 sRNAs 的失活会降低 S. typhimurium 对 GSNO 的抗性,并增加亚硝化蛋白的水平。这些结果促使我们评估这些 sRNAs 是否可能在抗硝化应激中发挥作用。RNA 谱分析被用作筛选来识别新的 RyhB-1 和 RyhB-2 调节的靶标。根据它们在硝化应激反应中的潜在作用及其在野生型、单突变和双突变菌株(ΔryhB1、ΔryhB2 和 ΔryhB1ΔryhB2)中的表达,通过定量 RT-PCR 分析了一组基因。在 GSNO 处理后,RyhB-1 和 RyhB-2 负调控编码细胞色素 bo 氧化酶(cyoABC)、细胞色素 bd 氧化酶(cydB)、细胞色素 b-562(cybC)的基因的表达,并正调控亚硝酸盐还原酶系统(nirBCD 操纵子)。综上所述,这些结果表明 RyhB-1 和 RyhB-2 可以精细地调节编码细胞色素氧化酶和硝酸盐还原酶系统的基因的表达,使细胞能够应对 GSNO 诱导的应激。