Dubrac Sarah, Touati Danièle
Institut Jacques Monod, CNRS-Universités Paris 6 et Paris 7, 2 place Jussieu, 75251 Paris Cedex 05, France.
Microbiology (Reading). 2002 Jan;148(Pt 1):147-56. doi: 10.1099/00221287-148-1-147.
Fur (ferric uptake regulation protein) activates sodB expression, increasing expression levels by a factor of seven and sodB transcript stability by a factor of three. Post-transcriptional regulation of sodB was investigated by searching for endoribonucleases that might be involved in sodB mRNA degradation. The activation of sodB expression was significantly reduced if both the RNaseE and RNaseIII genes were mutated. This correlated with cleavage at a palindromic sequence located in the 5' untranslated region of the sodB transcript. An RNA-binding assay showed that Fur did not directly protect the sodB transcript. It was hypothesized that the persistence of Fur-mediated activation of sodB expression in the RNase double mutant was probably due to an effect at the transcriptional level. Therefore, it was investigated whether Fur had a direct transcriptional effect in vitro. Fur bound the sodB promoter region with low affinity, but it was not able to increase sodB transcription. H-NS-mediated repression of sodB expression, which has been shown to be Fur-dependent, was characterized. No DNA-bending region was identified in the sodB promoter region. H-NS did not interfere with the post-transcriptional effect of Fur. Fur-dependent H-NS and the Fur post-transcriptional effect were not additive. This suggests that Fur and H-NS effects are indirect and may be mediated by a common intermediate.
铁摄取调节蛋白(Fur)激活sodB的表达,使表达水平提高7倍,sodB转录本稳定性提高3倍。通过寻找可能参与sodB mRNA降解的核糖核酸内切酶来研究sodB的转录后调控。如果RNaseE和RNaseIII基因都发生突变,sodB表达的激活会显著降低。这与sodB转录本5'非翻译区中一个回文序列处的切割有关。RNA结合试验表明,Fur不能直接保护sodB转录本。据推测,在RNase双突变体中Fur介导的sodB表达激活的持续性可能是由于转录水平的影响。因此,研究了Fur在体外是否有直接的转录作用。Fur以低亲和力结合sodB启动子区域,但不能增加sodB的转录。对已证明依赖于Fur的H-NS介导的sodB表达抑制进行了表征。在sodB启动子区域未鉴定到DNA弯曲区域。H-NS不干扰Fur的转录后作用。Fur依赖的H-NS和Fur的转录后作用不是相加的。这表明Fur和H-NS的作用是间接的,可能由一个共同的中间体介导。