Skouloubris S, Thiberge J M, Labigne A, De Reuse H
Unité de Pathogénie Bactérienne des Muqueuses, Institut Pasteur, 75724 Paris Cedex 15, France.
Infect Immun. 1998 Sep;66(9):4517-21. doi: 10.1128/IAI.66.9.4517-4521.1998.
We produced defined isogenic Helicobacter pylori ureI mutants to investigate the function of UreI, the product of one of the genes of the urease cluster. The insertion of a cat cassette had a strong polar effect on the expression of the downstream urease genes, resulting in very weak urease activity. Urease activity, measured in vitro, was normal in a strain in which ureI was almost completely deleted and replaced with a nonpolar cassette. In contrast to previous reports, we thus found that the product of ureI was not necessary for the synthesis of active urease. Experiments with the mouse-adapted H. pylori SS1 strain carrying the nonpolar ureI deletion showed that UreI is essential for H. pylori survival in vivo and/or colonization of the mouse stomach. The replacement of ureI with the nonpolar cassette strongly reduced H. pylori survival in acidic conditions (1-h incubation in phosphate-buffered saline solution at pH 2.2) in the presence of 10 mM urea. UreI is predicted to be an integral membrane protein and may therefore be involved in a transport process essential for H. pylori survival in vivo.
我们构建了明确的同基因幽门螺杆菌ureI突变体,以研究脲酶基因簇中一个基因的产物UreI的功能。cat盒的插入对下游脲酶基因的表达有强烈的极性效应,导致脲酶活性非常弱。在体外测量时,在一个ureI几乎完全缺失并用非极性盒替代的菌株中,脲酶活性正常。与之前的报道相反,我们因此发现ureI的产物对于活性脲酶的合成并非必需。用携带非极性ureI缺失的适应小鼠的幽门螺杆菌SS1菌株进行的实验表明,UreI对于幽门螺杆菌在体内的存活和/或在小鼠胃中的定殖至关重要。用非极性盒替代ureI在存在10 mM尿素的情况下,在酸性条件下(在pH 2.2的磷酸盐缓冲盐溶液中孵育1小时)强烈降低了幽门螺杆菌的存活率。UreI预计是一种整合膜蛋白,因此可能参与了幽门螺杆菌在体内存活所必需的转运过程。