Wadler Caryn S, Vanderpool Carin K
Department of Microbiology, University of Illinois at Urbana-Champaign, 601 South Goodwin Avenue, Urbana, IL 61801, USA.
Proc Natl Acad Sci U S A. 2007 Dec 18;104(51):20454-9. doi: 10.1073/pnas.0708102104. Epub 2007 Nov 27.
SgrS is a 227-nt small RNA that is expressed in Escherichia coli during glucose-phosphate stress, a condition associated with intracellular accumulation of glucose-6-phosphate caused by disruption of glycolytic flux. Under stress conditions, SgrS negatively regulates translation and stability of the ptsG mRNA, encoding the major glucose transporter, by means of a base pairing-dependent mechanism requiring the RNA chaperone Hfq. SgrS activity mitigates the effects of glucose-phosphate stress, and the present study has elucidated a function of SgrS that is proposed to contribute to the stress response. The 5' end of SgrS, upstream of the nucleotides involved in base pairing with the ptsG mRNA, contains a 43-aa ORF, sgrT, that is conserved in most species that contain SgrS-like small RNAs. The sgrT gene is translated in E. coli under conditions of glucose-phosphate stress. Analysis of alleles that separate the base pairing function of SgrS from the sgrT coding sequence revealed that either of these functions alone are sufficient for previously characterized SgrS phenotypes. SgrS-dependent down-regulation of ptsG mRNA stability does not require SgrT and SgrT by itself has no effect on ptsG mRNA stability. Cells expressing sgrT alone had a defect in glucose uptake even though they had nearly wild-type levels of PtsG (IICB(Glc)). Together, these data suggest that SgrS represents a previously unrecognized paradigm for small RNA (sRNA) regulators as a bifunctional RNA that encodes physiologically redundant but mechanistically distinct functions contributing to the same stress response.
SgrS是一种227个核苷酸的小RNA,在大肠杆菌受到葡萄糖-磷酸应激时表达,这种应激与糖酵解通量中断导致的6-磷酸葡萄糖在细胞内积累有关。在应激条件下,SgrS通过一种依赖碱基配对且需要RNA伴侣Hfq的机制,对编码主要葡萄糖转运蛋白的ptsG mRNA的翻译和稳定性进行负调控。SgrS的活性减轻了葡萄糖-磷酸应激的影响,本研究阐明了SgrS的一种功能,该功能被认为有助于应激反应。SgrS的5'端,即在与ptsG mRNA进行碱基配对的核苷酸上游,包含一个43个氨基酸的开放阅读框sgrT,在大多数含有SgrS样小RNA的物种中是保守的。sgrT基因在大肠杆菌中葡萄糖-磷酸应激条件下会被翻译。对将SgrS的碱基配对功能与sgrT编码序列分开的等位基因进行分析发现,单独的这些功能中的任何一个都足以产生先前已表征的SgrS表型。SgrS依赖的ptsG mRNA稳定性下调不需要SgrT,并且SgrT本身对ptsG mRNA稳定性没有影响。单独表达sgrT的细胞即使具有接近野生型水平的PtsG(IICB(Glc)),在葡萄糖摄取方面也存在缺陷。总之,这些数据表明SgrS代表了一种以前未被认识的小RNA(sRNA)调节因子模式,作为一种双功能RNA,它编码生理上冗余但机制上不同的功能,共同促成相同的应激反应。