Hryniewicz M, Sirko A, Pałucha A, Böck A, Hulanicka D
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw.
J Bacteriol. 1990 Jun;172(6):3358-66. doi: 10.1128/jb.172.6.3358-3366.1990.
The sequence of 1,973 nucleotides encompassing the region at and directly adjacent to the CysB-dependent promoter controlling expression and synthesis of the sulfate-thiosulfate transport system of Escherichia coli has been determined. The transcription start site has been mapped by primer extension. One open reading frame representing the first gene of the presumed sulfate transport operon was identified and designated cysP. The deduced amino acid sequence of the CysP polypeptide indicates the presence of a signal peptide. Expression of the cysP gene in the T7 promoter-polymerase system revealed the location of the gene product in the periplasm. Construction of a cysP insertional mutant and assays of binding and uptake of sulfate and thiosulfate by this mutant allowed the identification of the cysP gene product as a thiosulfate-binding protein. The TGA termination codon of cysP was found to overlap the putative ATG initiation codon of the next open reading frame, inferred as being essential for the sulfate transport system, and it was designated cysT. Preliminary sequence data from the corresponding region of the Salmonella typhimurium chromosome showed strictly homologous counterparts of the E. coli cysP and cysT genes.
已确定包含大肠杆菌硫酸盐 - 硫代硫酸盐转运系统表达和合成所依赖的CysB启动子及其紧邻区域的1973个核苷酸的序列。通过引物延伸确定了转录起始位点。鉴定出一个代表假定的硫酸盐转运操纵子第一个基因的开放阅读框,并将其命名为cysP。CysP多肽的推导氨基酸序列表明存在信号肽。cysP基因在T7启动子 - 聚合酶系统中的表达揭示了基因产物位于周质中。构建cysP插入突变体并对该突变体进行硫酸盐和硫代硫酸盐结合及摄取测定,从而鉴定出cysP基因产物为硫代硫酸盐结合蛋白。发现cysP的TGA终止密码子与下一个开放阅读框的假定ATG起始密码子重叠,推测该开放阅读框对硫酸盐转运系统至关重要,并将其命名为cysT。来自鼠伤寒沙门氏菌染色体相应区域的初步序列数据显示了大肠杆菌cysP和cysT基因的严格同源对应物。