Baños Rosa C, Pons José I, Madrid Cristina, Juárez Antonio
Institut de Bioenginyeria de Catalunya, Parc Científic de Barcelona, Edifici Hèlix. c/ Josep Samitier 1-5, 08028 Barcelona, Spain.
Departament de Microbiologia, Facultat de Biologia, Universitat de Barcelona, Avda Diagonal 645, 08028 Barcelona, Spain.
Microbiology (Reading). 2008 May;154(Pt 5):1281-1289. doi: 10.1099/mic.0.2007/015610-0.
The H-NS protein plays a significant role in the modulation of gene expression in Gram-negative bacteria. Whereas isolation and characterization of hns mutants in Escherichia coli, Salmonella and Shigella represented critical steps to gain insight into the modulatory role of H-NS, it has hitherto not been possible to isolate hns mutants in Yersinia. The hns mutation is considered to be deleterious in this genus. To study the modulatory role of H-NS in Yersinia we circumvented hns lethality by expressing in Y. enterocolitica a truncated H-NS protein known to exhibit anti-H-NS activity in E. coli (H-NST(EPEC)). Y. enterocolitica cells expressing H-NST(EPEC) showed an altered growth rate and several differences in the protein expression pattern, including the ProV protein, which is modulated by H-NS in other enteric bacteria. To further confirm that H-NST(EPEC) expression in Yersinia can be used to demonstrate H-NS-dependent regulation in this genus, we used this approach to show that H-NS modulates expression of the YmoA protein.
H-NS蛋白在革兰氏阴性菌基因表达的调控中发挥着重要作用。虽然在大肠杆菌、沙门氏菌和志贺氏菌中分离和鉴定hns突变体是深入了解H-NS调控作用的关键步骤,但迄今为止,尚无法在耶尔森氏菌中分离出hns突变体。hns突变在该菌属中被认为是有害的。为了研究H-NS在耶尔森氏菌中的调控作用,我们通过在小肠结肠炎耶尔森氏菌中表达一种已知在大肠杆菌中具有抗H-NS活性的截短型H-NS蛋白(H-NST(EPEC))来规避hns致死性。表达H-NST(EPEC)的小肠结肠炎耶尔森氏菌细胞显示出生长速率改变以及蛋白质表达模式的若干差异,包括ProV蛋白,该蛋白在其他肠道细菌中受H-NS调控。为了进一步证实耶尔森氏菌中H-NST(EPEC)的表达可用于证明该菌属中H-NS依赖性调控,我们采用此方法表明H-NS调节YmoA蛋白的表达。