Delor I, Kaeckenbeeck A, Wauters G, Cornelis G R
Unité de Microbiologie, Université Catholique de Louvain, Brussels, Belgium.
Infect Immun. 1990 Sep;58(9):2983-8. doi: 10.1128/iai.58.9.2983-2988.1990.
The gene encoding the heat-stable enterotoxin (yst) was cloned from the chromosome of Yersinia enterocolitica W1024 (serotype O:9), and the nucleotide sequence was determined. The yst gene encodes a 71-amino-acid polypeptide. The C-terminal 30 amino acids of the predicted protein exactly correspond to the amino acid sequence of the toxin extracted from culture supernatants (T. Takao, N. Tominaga, and Y. Shimonishi, Biochem. Biophys. Res. Commun. 125:845-851, 1984). The N-terminal 18 amino acids have the properties of a signal sequence. The central 22 residues are removed during or after the secretion process. This organization in three domains (Pre, Pro, and mature Yst) resembles that of the enterotoxin STa of Escherichia coli. The degree of conservation between the E. coli and Y. enterocolitica toxins is much lower in the Pre and the Pro domains than in the mature proteins. The mature toxin of Y. enterocolitica is much larger than that of E. coli, but the active domain appears to be highly conserved. The yst gene of Y. enterocolitica introduced in E. coli K-12 directed the secretion of an active toxin. The cloned yst gene was used as an epidemiological probe among a collection of 174 strains representative of all Yersinia species except Yersinia pestis and numerous Y. enterocolitica subgroups. In Y. enterocolitica, there was a clear-cut difference between pathogenic and nonpathogenic strains: 89 of 89 pathogenic and none of 51 nonpathogenic strains contained yst-homologous DNA, suggesting that Yst is involved in pathogenesis. Among the other Yersinia species, only four strains of Yersinia kristensenii had DNA homologous to yst.
编码热稳定肠毒素(yst)的基因从小肠结肠炎耶尔森菌W1024(血清型O:9)的染色体中克隆出来,并测定了核苷酸序列。yst基因编码一种71个氨基酸的多肽。预测蛋白的C末端30个氨基酸与从培养上清液中提取的毒素的氨基酸序列完全一致(T. 高尾、N. 富永、Y. 下西,《生物化学与生物物理学研究通讯》125:845 - 851,1984)。N末端18个氨基酸具有信号序列的特性。中间的22个残基在分泌过程中或分泌后被去除。这种三个结构域(前体、原毒素和成熟Yst)的结构类似于大肠杆菌的肠毒素STa。大肠杆菌和小肠结肠炎耶尔森菌毒素在前体和原毒素结构域的保守程度比成熟蛋白低得多。小肠结肠炎耶尔森菌的成熟毒素比大肠杆菌的大得多,但活性结构域似乎高度保守。导入大肠杆菌K - 12中的小肠结肠炎耶尔森菌的yst基因指导了活性毒素的分泌。克隆的yst基因被用作一种流行病学探针,用于检测174株代表除鼠疫耶尔森菌外的所有耶尔森菌属及众多小肠结肠炎耶尔森菌亚群的菌株。在小肠结肠炎耶尔森菌中,致病菌株和非致病菌株之间存在明显差异:89株致病菌株中有89株含有yst同源DNA,而51株非致病菌株中无一含有,这表明Yst与致病机制有关。在其他耶尔森菌属中,只有4株克氏耶尔森菌的DNA与yst同源。