Chen N Y, Zhang J J, Paulus H
Department of Metabolic Regulation, Boston Biomedical Research Institute, Massachusetts 02114.
J Gen Microbiol. 1989 Nov;135(11):2931-40. doi: 10.1099/00221287-135-11-2931.
The aspartokinase II (ask) operon of Bacillus subtilis consists of two in-phase overlapping genes that encode the two subunits of the lysine-sensitive isoenzyme of aspartokinase (ATP:L-aspartate 4-phosphotransferase, EC 2.7.2.4). Transduction mapping of the ask operon, inactivated by recombinational insertion of a cat marker, indicates a chromosomal location (about 253 degrees) between leuA and aroG. ask is thus remote from aecB, eliminating aecB as a possible locus for the structural gene of aspartokinase II, but close to aecA and uvrB. The nucleotide sequence of a 2 kb DNA fragment just upstream of the ask operon was determined and found to contain two open reading frames. The deduced amino acid sequence of the distal reading frame exhibits extensive homology with Escherichia coli thioredoxin and that of the proximal one, which overlaps with the ask promoter, is homologous to the deduced product of the E. coli uvrC gene. Insertional mutagenesis of the proximal open reading frame led to a mitomycin-sensitive phenotype, consistent with a role in DNA repair. In conjunction with the data of M. Petricek, L. Rutberg & L. Hederstedt [FEMS Microbiology Letters 61, 85-88] our results define the nucleotide sequence of an 8.8 kb segment of the B. subtilis chromosome near 253 degrees and the following order of genes: trx-uvrB-ask-orfX-sdhC-sdhA-sdhB-orfY++ +-gerE.
枯草芽孢杆菌的天冬氨酸激酶II(ask)操纵子由两个同相位重叠基因组成,这两个基因编码天冬氨酸激酶(ATP:L-天冬氨酸4-磷酸转移酶,EC 2.7.2.4)的赖氨酸敏感同工酶的两个亚基。通过cat标记的重组插入而失活的ask操纵子的转导作图表明,其在染色体上的位置(约253度)位于leuA和aroG之间。因此,ask远离aecB,排除了aecB作为天冬氨酸激酶II结构基因的可能位点,但靠近aecA和uvrB。测定了ask操纵子上游刚好一个2 kb DNA片段的核苷酸序列,发现其包含两个开放阅读框。远端阅读框推导的氨基酸序列与大肠杆菌硫氧还蛋白具有广泛的同源性,而与ask启动子重叠的近端阅读框推导的氨基酸序列与大肠杆菌uvrC基因的推导产物同源。近端开放阅读框的插入诱变导致对丝裂霉素敏感的表型,这与在DNA修复中的作用一致。结合M. Petricek、L. Rutberg和L. Hederstedt [FEMS微生物学快报61,85 - 88]的数据,我们的结果确定了枯草芽孢杆菌染色体253度附近一个8.8 kb片段的核苷酸序列以及以下基因顺序:trx - uvrB - ask - orfX - sdhC - sdhA - sdhB - orfY++ +-gerE。