Sakanyan Vehary, Petrosyan Pavel, Lecocq Michèle, Boyen Anne, Legrain Christiane, Demarez Marc, Hallet Jean-Noël, Glansdorff Nicolas
Laboratoire de Biotechnologie, Faculté des Sciences et des Techniques, Université de Nantes, 2, rue de la Houssinière, F-44072 Nantes Cedex 03, France.
Pharmagen, Knunyants Street 4, Yerevan 375010, Republic of Armenia.
Microbiology (Reading). 1996 Jan;142 ( Pt 1):99-108. doi: 10.1099/13500872-142-1-99.
A cluster of arginine biosynthetic genes of Corynebacterium glutamicum ATCC 13032, comprising argJ, argB and argD as well as part of argC and argF, has been cloned by heterologous complementation of an Escherichia coli argE mutant. The gene order has been established as argCJBDF by sequencing the entire 4.4 kb cloned DNA fragment. The C. glutamicum argB gene can be transcribed in E. coli cells from an internal promoter located in the coding part of the preceding argJ gene, whereas transcription of the argJ gene appears vector-dependent. Expression of the corynebacterial argB gene is repressed by arginine in the native host but not in recombinant E. coli cells. Feedback inhibition of the corresponding N-acetylglutamate kinase activity was observed both in cell extracts of C. glutamicum and in recombinant E. coli argB auxotrophic strains. Extracts of E. coli cells carrying cloned corynebacterial DNA display an ornithine acetyltransferase activity (encoded by argJ) which alleviates the acetylornithinase (encoded by argE) deficiency of the enterobacterial host. In contrast to Bacillus stearothermophilus ornithine acetyltransferase which also exhibits acetylglutamate synthase activity, C. glutamicum ornithine acetyltransferase appears monofunctional. ArgA and ArgB proteins from different sources share highly significant similarities. The evolutionary implications of these data are discussed.
通过对大肠杆菌argE突变体的异源互补,克隆了谷氨酸棒杆菌ATCC 13032的一组精氨酸生物合成基因,包括argJ、argB和argD以及部分argC和argF。通过对整个4.4 kb克隆DNA片段进行测序,确定基因顺序为argCJBDF。谷氨酸棒杆菌argB基因可在大肠杆菌细胞中从位于前一个argJ基因编码区的内部启动子转录,而argJ基因的转录似乎依赖于载体。在天然宿主中,精氨酸可抑制棒杆菌argB基因的表达,但在重组大肠杆菌细胞中则不会。在谷氨酸棒杆菌的细胞提取物和重组大肠杆菌argB营养缺陷型菌株中均观察到了对相应N-乙酰谷氨酸激酶活性的反馈抑制。携带克隆的棒杆菌DNA的大肠杆菌细胞提取物表现出鸟氨酸乙酰转移酶活性(由argJ编码),可缓解肠道细菌宿主的乙酰鸟氨酸酶(由argE编码)缺陷。与也具有乙酰谷氨酸合酶活性的嗜热脂肪芽孢杆菌鸟氨酸乙酰转移酶不同,谷氨酸棒杆菌鸟氨酸乙酰转移酶似乎是单功能的。来自不同来源的ArgA和ArgB蛋白具有高度显著的相似性。讨论了这些数据的进化意义。