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3′ Untranslated Region-Dependent Degradation of the aceA mRNA, Encoding the Glyoxylate Cycle Enzyme Isocitrate Lyase, by RNase E/G in Corynebacterium glutamicum

机译:谷氨酸棒杆菌中RNase E / G对编码乙醛酸循环酶异柠檬酸裂解酶的aceA mRNA的3'非翻译区依赖性降解

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We previously reported that the Corynebacterium glutamicum RNase E/G encoded by the rneG gene (NCgl2281) is required for the 5′ maturation of 5S rRNA. In the search for the intracellular target RNAs of RNase E/G other than the 5S rRNA precursor, we detected that the amount of isocitrate lyase, an enzyme of the glyoxylate cycle, increased in rneG knockout mutant cells grown on sodium acetate as the sole carbon source. Rifampin chase experiments showed that the half-life of the aceA mRNA was about 4 times longer in the rneG knockout mutant than in the wild type. Quantitative real-time PCR analysis also confirmed that the level of aceA mRNA was approximately 3-fold higher in the rneG knockout mutant strain than in the wild type. Such differences were not observed in other mRNAs encoding enzymes involved in acetate metabolism. Analysis by 3′ rapid amplification of cDNA ends suggested that RNase E/G cleaves the aceA mRNA at a single-stranded AU-rich region in the 3′ untranslated region (3′-UTR). The lacZ fusion assay showed that the 3′-UTR rendered lacZ mRNA RNase E/G dependent. These findings indicate that RNase E/G is a novel regulator of the glyoxylate cycle in C. glutamicum .
机译:我们之前曾报道过,rneG基因(NCgl2281)编码的谷氨酸棒杆菌RNase E / G是5S rRNA 5'成熟所必需的。在寻找5S rRNA前体以外的RNase E / G的细胞内靶RNA时,我们发现异柠檬酸裂解酶(乙醛酸循环酶)的量在以乙酸钠为唯一碳的rneG敲除突变体细胞中增加了。资源。利福平追踪实验表明,rneG基因敲除突变体中aceA mRNA的半衰期比野生型长约4倍。实时定量PCR分析还证实,rneG基因敲除突变株中aceA mRNA的水平比野生型高约3倍。在其他参与乙酸酯代谢的编码酶的mRNA中未观察到这种差异。通过3'cDNA末端快速扩增的分析表明,RNase E / G在3'非翻译区(3'-UTR)的单链富AU区域切割aceA mRNA。 lacZ融合试验表明3'-UTR使得lacZ mRNA RNase E / G依赖性。这些发现表明,RNase E / G是谷氨酸棒杆菌中乙醛酸循环的新型调节剂。

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