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铜绿假单胞菌磷酸盐特异性转运(pst)操纵子的分子分析。

Molecular analysis of the phosphate-specific transport (pst) operon of Pseudomonas aeruginosa.

作者信息

Nikata T, Sakai Y, Shibat K, Kato J, Kuroda A, Ohtake H

机构信息

Department of Fermentation Technology, Hiroshima University, Japan.

出版信息

Mol Gen Genet. 1996 Apr 10;250(6):692-8. doi: 10.1007/BF02172980.

Abstract

The organization of the phosphate-specific transport (pst) operon in Pseudomonas aeruginosa has been determined. The gene order of the pst operon is pstC, pstA, pstB, phoU, and a well-conserved Pho box sequence (16/18 bases identical) exists in the promoter region. The most striking difference from the known Escherichia coli pst operon is the lack of the pstS gene encoding a periplasmic phosphate (Pi)-binding protein. Even though the three pst genes were absolutely required for P(i)-specific transport, expression of the pst operon at high levels did not increase P(i) uptake in P. aeruginosa. DNA sequences for the pstB and phoU genes have been determined previously. The newly identified pstC and pstA genes encode possible integral membrane proteins of 677 amino acids (M(r) 73,844) and 513 amino acids (M(r) 56,394) respectively. The amino acid sequences of PstC and PstA predict that these proteins contain a long hydrophilic domain not seen in their E. coli counterparts. A chromosomal deletion of the entire pst operon rendered P. aeruginosa unable to repress P(i) taxis under conditions of P(i) excess. The phoU and pstB genes are essential for repressing P(i) taxis. However, mutants lacking either PstC or PstA alone were able to repress P(i) taxis under conditions of P(i) excess.

摘要

已确定铜绿假单胞菌中磷酸特异性转运(pst)操纵子的组织形式。pst操纵子的基因顺序为pstC、pstA、pstB、phoU,并且在启动子区域存在一个保守性良好的Pho盒序列(16/18个碱基相同)。与已知的大肠杆菌pst操纵子最显著的差异是缺少编码周质磷酸(Pi)结合蛋白的pstS基因。尽管这三个pst基因对于Pi特异性转运是绝对必需的,但pst操纵子的高水平表达并未增加铜绿假单胞菌对Pi的摄取。pstB和phoU基因的DNA序列先前已确定。新鉴定的pstC和pstA基因分别编码可能的整合膜蛋白,其含有677个氨基酸(Mr 73,844)和513个氨基酸(Mr 56,394)。PstC和PstA的氨基酸序列预测这些蛋白含有一个在其大肠杆菌对应物中未见的长亲水区。整个pst操纵子的染色体缺失使铜绿假单胞菌在Pi过量条件下无法抑制Pi趋化性。phoU和pstB基因对于抑制Pi趋化性至关重要。然而,单独缺失PstC或PstA的突变体在Pi过量条件下能够抑制Pi趋化性。

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