Gamper M, Zimmermann A, Haas D
Mikrobiologisches Institut, Eidgenössische Technische Hochschule, CH-8092 Zürich, Switzerland.
J Bacteriol. 1991 Aug;173(15):4742-50. doi: 10.1128/jb.173.15.4742-4750.1991.
The arcDABC operon of Pseudomonas aeruginosa encodes the enzymes of the arginine deiminase pathway, which is inducible under conditions of oxygen limitation and serves to generate ATP from arginine. The 5' end of arc mRNA extracted from anaerobically grown cells was determined by S1 and primer extension mapping. The transcription initiation site was located upstream of the arcD gene and 41.5 bp downstream of the center of the sequence TTGAC....ATCAG. This sequence, termed the ANR box, is similar to the consensus FNR recognition site of Escherichia coli. Transcription of the arc operon in P. aeruginosa was strongly decreased by a deletion of the TTGAC half site or by a mutation in the anr gene, which is known to code for the FNR-like regulatory protein ANR. During a transition from aerobic to anaerobic growth conditions, the concentrations of arc mRNAs and the levels of the ArcD and ArcA proteins rose in a parallel fashion. Mutational analysis of the arc promoter region led to the conclusion that the distance between the ANR box and the -10 promoter region is important for promoter strength, whereas the -35 region does not appear to be critical for arc promoter function. These findings and previous results indicate that anaerobic induction of the arc operon occurs at the level of transcription and requires the ANR box in cis and the ANR protein in trans.
铜绿假单胞菌的arcDABC操纵子编码精氨酸脱亚氨酶途径的酶,该途径在氧气限制条件下可被诱导,并用于从精氨酸生成ATP。通过S1和引物延伸图谱确定了从厌氧生长细胞中提取的arc mRNA的5'末端。转录起始位点位于arcD基因上游,在序列TTGAC....ATCAG中心下游41.5 bp处。该序列称为ANR框,与大肠杆菌的共有FNR识别位点相似。通过缺失TTGAC半位点或通过anr基因中的突变(已知该基因编码FNR样调节蛋白ANR),铜绿假单胞菌中arc操纵子的转录强烈降低。在从需氧生长条件转变为厌氧生长条件期间,arc mRNA的浓度以及ArcD和ArcA蛋白的水平以平行方式上升。对arc启动子区域的突变分析得出结论,ANR框与-10启动子区域之间的距离对启动子强度很重要,而-35区域似乎对arc启动子功能并不关键。这些发现和先前的结果表明,arc操纵子的厌氧诱导发生在转录水平,并且在顺式中需要ANR框,在反式中需要ANR蛋白。