Schlictman D, Shankar S, Chakrabarty A M
Department of Microbiology and Immunology, College of Medicine, University of Illinois, Chicago 60612, USA.
Mol Microbiol. 1995 Apr;16(2):309-20. doi: 10.1111/j.1365-2958.1995.tb02303.x.
The algR2 (also known as algQ) gene of Pseudomonas aeruginosa has previously been identified as being necessary for alginate production at 37 degrees C. We have cloned two genes, from a cosmid library of Escherichia coli, which can restore mucoidy to an algR2 mutant of P. aeruginosa. The complementing regions of both cosmids were localized by subcloning restriction fragments. One of the E. coli genes identified here has not previously been described; we have named this gene rnk (regulator of nucleoside diphosphate kinase). It encodes a 14.9 kDa protein with no homology to any other protein. The other gene, sspA, is a regulator involved in stationary-phase regulation in E. coli. Either gene will restore mucoidy to an algR2-deficient strain of P. aeruginosa. AlgR2 has been shown to regulate at least two enzymes, succinyl-CoA synthetase (Scs) and nucleoside diphosphate kinase (Ndk), which form a complex in P. aeruginosa. When we examined the ability of the E. coli analogues to regulate Ndk, we found that rnk but not sspA was able to restore Ndk activity to the P. aeruginosa algR2 mutant. Furthermore, rnk was able to restore growth of the algR2 mutant in the presence of Tween 20, which inhibits other Ndk-like activities.
铜绿假单胞菌的algR2(也称为algQ)基因先前已被确定为在37摄氏度下产生藻酸盐所必需的基因。我们从大肠杆菌的黏粒文库中克隆了两个基因,它们可使铜绿假单胞菌的algR2突变体恢复黏液状。通过亚克隆限制片段对两个黏粒的互补区域进行了定位。此处鉴定出的一个大肠杆菌基因以前未曾描述过;我们将此基因命名为rnk(核苷二磷酸激酶调节因子)。它编码一种14.9 kDa的蛋白质,与任何其他蛋白质均无同源性。另一个基因sspA是参与大肠杆菌稳定期调节的调节因子。任一基因均可使铜绿假单胞菌的algR2缺陷菌株恢复黏液状。已证明AlgR2可调节至少两种酶,琥珀酰辅酶A合成酶(Scs)和核苷二磷酸激酶(Ndk),它们在铜绿假单胞菌中形成复合物。当我们检测大肠杆菌类似物调节Ndk的能力时,我们发现rnk而非sspA能够使铜绿假单胞菌algR2突变体恢复Ndk活性。此外,rnk能够在吐温20存在的情况下恢复algR2突变体的生长,吐温20可抑制其他类似Ndk的活性。