van der Ploeg J R, Weiss M A, Saller E, Nashimoto H, Saito N, Kertesz M A, Leisinger T
Mikrobiologisches Institut, Swiss Federal Institute of Technology, ETH-Zentrum, Zürich, Switzerland.
J Bacteriol. 1996 Sep;178(18):5438-46. doi: 10.1128/jb.178.18.5438-5446.1996.
Genes whose expression is regulated by sulfate starvation in Escherichia coli were identified by generating random translational lacZ fusions in the chromosome with the lambda placMu9 system. Nine lacZ fusion strains which expressed beta-galactosidase after growth under sulfate starvation conditions but not after growth in the presence of sulfate were found. These included two strains with insertions in the dmsA and rhsD genes, respectively, and seven strains in which the insertions were located within a 1.8-kb region downstream of hemB at 8.5 minutes on the E. coli chromosome. Analysis of the nucleotide sequence of this region indicated the presence of four open reading frames designated tauABCD. Disruption of these genes resulted in the loss of the ability to utilize taurine (2-aminoethanesulfonate) as a source of sulfur but did not affect the utilization of a range of other aliphatic sulfonates as sulfur sources. The TauA protein contained a putative signal peptide for transport into the periplasm; the TauB and TauC proteins showed sequence similarity to ATP-binding proteins and membrane proteins, respectively, of ABC-type transport systems; and the TauD protein was related in sequence to a dichlorophenoxyacetic acid dioxygenase. We therefore suggest that the proteins encoded by tauABC constitute an uptake system for taurine and that the product of tauD is involved in the oxygenolytic release of sulfite from taurine. The transcription initiation site was detected 26 to 27 bp upstream of the translational start site of tauA. Expression of the tauD gene was dependent on CysB, the transcriptional activator of the cysteine regulon.
利用λplacMu9系统在大肠杆菌染色体中产生随机翻译型lacZ融合,从而鉴定出其表达受硫酸盐饥饿调控的基因。发现了9个lacZ融合菌株,这些菌株在硫酸盐饥饿条件下生长后能表达β-半乳糖苷酶,但在有硫酸盐存在的情况下生长后则不能。其中包括分别在dmsA和rhsD基因中插入的两个菌株,以及7个插入位于大肠杆菌染色体8.5分钟处hemB下游1.8 kb区域内的菌株。对该区域核苷酸序列的分析表明存在四个开放阅读框,命名为tauABCD。这些基因的破坏导致利用牛磺酸(2-氨基乙磺酸盐)作为硫源的能力丧失,但不影响利用一系列其他脂肪族磺酸盐作为硫源。TauA蛋白含有一个推测的用于转运到周质中的信号肽;TauB和TauC蛋白分别与ABC型转运系统的ATP结合蛋白和膜蛋白显示出序列相似性;TauD蛋白在序列上与二氯苯氧基乙酸双加氧酶相关。因此,我们认为tauABC编码的蛋白构成了一个牛磺酸摄取系统,并且tauD的产物参与了从牛磺酸中氧解释放亚硫酸盐的过程。转录起始位点在tauA翻译起始位点上游26至27 bp处被检测到。tauD基因的表达依赖于CysB,即半胱氨酸调节子的转录激活因子。