Piechaczek K, Dobrindt U, Schierhorn A, Fischer G S, Hecker M, Hacker J
Institut für Molekulare Infektionsbiologie, Würzburg, Germany.
Int J Med Microbiol. 2000 Mar;290(1):75-84. doi: 10.1016/S1438-4221(00)80110-1.
The uropathogenic Escherichia coli strain 536 (O6:K15:H31) carries four distinct DNA regions in its chromosome, termed pathogenicity islands (PAIs I536 to IV536). Each of these PAIs encodes at least one virulence factor. All four PAIs are associated with tRNA genes. PAI I536 and PAI II536 can be spontaneously deleted from the chromosome by homologous recombination between flanking direct repeats. The deletion of PAI II536 results in the truncation of the associated gene leuX encoding the tRNALeu. This tRNA influences the expression of various virulence traits. In order to get a deeper insight into the role of PAI I536/II536 and of the tRNA5LeU for the protein expression, the protein expression patterns of Escherichia coli 536 and different derivatives were studied. Differences in the protein expression patterns of the wild-type strain Escherichia coli 536, its mutants 536-21 (PAI I536-, PAI II536-, leuX-), 536delta102 (PAI I536+, PAI II536+, leuX-) as well as of the strain 536R3 (PAI I536-, PAI II536-, leuX+) were analyzed by two-dimensional polyacrylamide gel electrophoresis and MALDI-TOF mass spectrometry. We identified about 39 different intracellular proteins whose expression is markedly altered in the different strain backgrounds. These differences can be linked either to the presence or absence of the PAI I536 and PAI II536 or to that of the tRNA gene leuX. The identities of 34 proteins have been determined by MALDI-TOF-MS. The identification of five proteins was not possible. The results suggest that proteome analysis is an efficient approach to study differences in global gene expression. The comparison of protein expression patterns of the uropathogenic E. coli strain 536 and different derivatives revealed that in this strain the expression of various proteins including those encoded by many housekeeping genes is affected by the presence of PAI I536 and Pai II536 or by that of the tRNA5Leu.
尿路致病性大肠杆菌菌株536(O6:K15:H31)在其染色体中携带四个不同的DNA区域,称为致病岛(PAI I536至IV536)。这些致病岛中的每一个都编码至少一种毒力因子。所有四个致病岛都与tRNA基因相关。PAI I536和PAI II536可通过侧翼同向重复序列之间的同源重组从染色体上自发缺失。PAI II536的缺失导致编码tRNALeu的相关基因leuX截短。这种tRNA影响各种毒力性状的表达。为了更深入了解PAI I536/II536和tRNA5LeU在蛋白质表达中的作用,研究了大肠杆菌536及其不同衍生物的蛋白质表达模式。通过二维聚丙烯酰胺凝胶电泳和基质辅助激光解吸电离飞行时间质谱分析了野生型大肠杆菌536菌株、其突变体536-21(PAI I536-、PAI II536-、leuX-)、536delta102(PAI I536+、PAI II536+、leuX-)以及菌株536R3(PAI I536-、PAI II536-、leuX+)的蛋白质表达模式差异。我们鉴定出约39种不同的细胞内蛋白质,其表达在不同菌株背景下有明显改变。这些差异可能与PAI I536和PAI II536的存在与否或tRNA基因leuX的存在与否有关。34种蛋白质的身份已通过基质辅助激光解吸电离飞行时间质谱法确定。无法鉴定出五种蛋白质。结果表明蛋白质组分析是研究全局基因表达差异的有效方法。尿路致病性大肠杆菌菌株536及其不同衍生物的蛋白质表达模式比较表明,在该菌株中,包括许多管家基因编码的蛋白质在内的各种蛋白质的表达受PAI I536和PAI II536或tRNA5Leu的存在影响。