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Transcriptional regulation of the aconitase genes (acnA and acnB) of Escherichia coli

机译:大肠杆菌的Aconitase基因(ACNA和ACNB)的转录调节

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Escherichia colicontains two differentially regulated aconitase genes, acnA and acnB. Two acnA promoters transcribing from start points located 407 bp (P1acnA) and 50 bp (P2acnA) upstream of the acnA coding region, and one acnB promoter (PacnB) with a start point 95 bp upstream of the acnB coding region, were identified by primer extension analysis. A 2.8 kb acnA monocistronic transcript was detected by Northern blot hybridization, but only in redox-stressed (methyl-viologen-treated) cultures, and a 2.5 kb acnB monocistronic transcript was detected in exponential- but not stationary-phase cultures. These findings are consistent with previous observations that acnA is specifically subject to SoxRS-mediated activation, whereas acnB encodes the major aconitase that is synthesized earlier in the growth cycle than AcnA. Further studies with acn-lacZ gene fusions and a wider range of transcription regulators indicated that acnA expression is initiated by σ38 from P1acnA’ and from P2acnA it is activated directly or indirectly by CRP, FruR, Fur and SoxRS, anc repressed by ArcA and FNR. In contrast, acnB expression is activated by CRP and repressed by ArcA, FruR and Fis from PacnB Comparable studies with fum-lacZ fusions indicated that transcription of fumC, but not of fumA or fumB, is initiated by RNA polymerase containing σ38. It is concluded that AcnB is the major citric acid cycle enzyme, whereas AcnA is an aerobic stationary-phase enzyme that is specifically induced by iron and redox-stress.
机译:大肠杆菌肠道两种差异调节的穴位酶基因,ACNA和ACNB。通过引物鉴定了从ACNA编码区域的上游的407bp(p1acna)和50bp(p2acna)和50bp(p2acna)的起始点转录的两种ACNA启动子,以及ACNB促进剂(PACNB)上游的ACNB编码区域的起点95bp扩展分析。通过Northern印迹杂交检测2.8kb ACNA单闭转录物,但仅在氧化还原(甲基 - viologen处理)培养物中,并在指数 - 但不是稳定性相培养物中检测到2.5kb的acnb单闭转录物。这些发现与先前的观察结果一致,即ACNA特异性受到SOXR介导的活化的影响,而ACNB编码在比ACNA的生长循环中以前合成的主要遮阳性酶。通过ACN-LacZ基因融合和更广泛的转录调节剂进一步研究表明,ACNA表达由P1AcNA'的σ38引发,并且来自P2Acna,它通过CRP,Frur,毛皮和Soxrs直接或间接地活化,Acca和FNR压抑。相反,通过CRP激活ACNB表达,并通过ARCA,FIS和FIS从PACNB与FUM-LACZ融合的比较研究表明,含有σ38的RNA聚合酶引发FUMC但不具有福音或烟雾的转录。结论是,ACNB是主要柠檬酸循环酶,而ACNA是一种有氧固定相酶,由铁和氧化还原胁迫具体诱导。

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