Laboratorio de Microbiología, Facultad de Ciencias Biológicas y Facultad de Medicina, Universidad Andres Bello, República 217, Santiago de Chile, Chile.
Res Microbiol. 2009 Dec;160(10):800-8. doi: 10.1016/j.resmic.2009.09.016. Epub 2009 Oct 14.
A novel pathogenicity island, SPI-18, carries the taiA-hlyE operon, encoding virulence factors in Salmonella Typhi. To determine the effects of certain environmental conditions on the expression of these genes, beta-galactosidase assays, RT-PCR reactions, western blot analyses and measurement of hemolytic activity were performed. The conditions studied are those likely found by S. Typhi during infection in the human host. We found RpoS-dependent transcriptional upregulation in low pH and high osmolarity for both genes. Our results show that oxygen depletion apparently did not affect transcription of the taiA-hlyE operon. On the other hand, the transcriptional regulator Crp, previously described as an activator of hlyE transcription in Escherichia coli, is involved in transcriptional repression of hlyE in S. Typhi. Moreover, addition of glucose to the growth medium results in decreasing the hlyE mRNA, suggesting that there is another factor related to catabolite repression different from Crp and involved in downregulation of hlyE in S. Typhi.
一个新的致病性岛 SPI-18 携带 taiA-hlyE 操纵子,编码伤寒沙门氏菌的毒力因子。为了确定某些环境条件对这些基因表达的影响,进行了β-半乳糖苷酶测定、RT-PCR 反应、western blot 分析和溶血活性测定。研究的条件是伤寒沙门氏菌在人体感染过程中可能遇到的条件。我们发现,这两个基因在低 pH 和高渗透压下都依赖于 RpoS 进行转录上调。我们的结果表明,缺氧显然不会影响 taiA-hlyE 操纵子的转录。另一方面,先前在大肠杆菌中描述为 hlyE 转录激活剂的转录调节因子 Crp,参与伤寒沙门氏菌中 hlyE 的转录抑制。此外,在生长培养基中添加葡萄糖会导致 hlyE mRNA 减少,表明存在另一种与分解代谢物阻遏有关的因子,不同于 Crp,并参与伤寒沙门氏菌中 hlyE 的下调。