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An Extracytoplasmic Function Sigma Factor Acts as a General Stress Response Regulator in Sinorhizobium meliloti▿

机译:胞外功能西格玛因子充当melinohizobiummeliloti▿中的一般应激反应调节剂。

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摘要

Sinorhizobium meliloti genes transcriptionally up-regulated after heat stress, as well as upon entry into stationary phase, were identified by microarray analyses. Sixty stress response genes were thus found to be up-regulated under both conditions. One of them, rpoE2 (smc01506), encodes a putative extracytoplasmic function (ECF) sigma factor. We showed that this sigma factor controls its own transcription and is activated by various stress conditions, including heat and salt, as well as entry into stationary phase after either carbon or nitrogen starvation. We also present evidence that the product of the gene cotranscribed with rpoE2 negatively regulates RpoE2 activity, and we therefore propose that it plays the function of anti-sigma factor. By combining transcriptomic, bioinformatic, and quantitative reverse transcription-PCR analyses, we identified 44 RpoE2-controlled genes and predicted the number of RpoE2 targets to be higher. Strikingly, more than one-third of the 60 stress response genes identified in this study are RpoE2 targets. Interestingly, two genes encoding proteins with known functions in stress responses, namely, katC and rpoH2, as well as a second ECF-encoding gene, rpoE5, were found to be RpoE2 regulated. Altogether, these data suggest that RpoE2 is a major global regulator of the general stress response in S. meliloti. Despite these observations, and although this sigma factor is well conserved among alphaproteobacteria, no in vitro nor in planta phenotypic difference from the wild-type strain could be detected for rpoE2 mutants. This therefore suggests that other important actors in the general stress response have still to be identified in S. meliloti.
机译:通过微阵列分析鉴定了热应激后以及进入固定相后转录上调的中华根瘤菌基因。因此发现在两种条件下都有60个应激反应基因被上调。其中之一rpoE2(smc01506)编码一个假定的胞浆外功能(ECF)sigma因子。我们表明,该sigma因子控制其自身的转录,并被各种应激条件激活,包括热和盐,以及碳或氮饥饿后进入固定相。我们还提供了证据,证明与rpoE2共转录的基因产物负调控RpoE2活性,因此我们建议其发挥抗sigma因子的功能。通过结合转录组学,生物信息学和定量逆转录-PCR分析,我们确定了44个RpoE2控制的基因,并预测RpoE2靶标的数量会更高。令人惊讶的是,在这项研究中确定的60个应激反应基因中,有超过三分之一是RpoE2靶标。有趣的是,发现两个编码在应激反应中具有已知功能的蛋白质的基因,即katC和rpoH2,以及第二个ECF编码基因,rpoE5,都受到RpoE2调控。总之,这些数据表明RpoE2是苜蓿链球菌总体胁迫反应的主要全球调节因子。尽管有这些观察结果,并且尽管该sigma因子在α变形杆菌中是非常保守的,但是rpoE2突变体在体外和植物中均未检测到与野生型菌株的表型差异。因此,这表明在沙门氏菌中仍需确定总体应激反应中的其他重要因素。

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