Diep Duong Thi Hong, Phuong Nguyen Thi Thanh, Hlaing Mya Myintzu, Srimanote Potjanee, Tungpradabkul Sumalee
Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
Department of Clinical Microbiology, Faculty of Medical Technology, Mahidol University, Salaya 73170, Thailand.
Int J Bacteriol. 2015;2015:623967. doi: 10.1155/2015/623967. Epub 2015 Dec 22.
Burkholderia pseudomallei is the causative agent of melioidosis. The complete genome sequences of this pathogen have been revealed, which explain some pathogenic mechanisms. In various hostile conditions, for example, during nitrogen and amino acid starvation, bacteria can utilize alternative sigma factors such as RpoS and RpoN to modulate genes expression for their adaptation and survival. In this study, we demonstrate that mutagenesis of rpoN2, which lies on chromosome 2 of B. pseudomallei and encodes a homologue of the sigma factor RpoN, did not alter nitrogen and amino acid utilization of the bacterium. However, introduction of B. pseudomallei rpoN2 into E. coli strain deficient for rpoN restored the ability to utilize amino acids. Moreover, comparative partial proteomic analysis of the B. pseudomallei wild type and its rpoN2 isogenic mutant was performed to elucidate its amino acids utilization property which was comparable to its function found in the complementation assay. By contrast, the rpoN2 mutant exhibited decreased katE expression at the transcriptional and translational levels. Our finding indicates that B. pseudomallei RpoN2 is involved in a specific function in the regulation of catalase E expression.
类鼻疽伯克霍尔德菌是类鼻疽的病原体。该病原体的完整基因组序列已被揭示,这解释了一些致病机制。在各种恶劣条件下,例如在氮和氨基酸饥饿期间,细菌可以利用替代的σ因子,如RpoS和RpoN来调节基因表达以实现自身适应和存活。在本研究中,我们证明了位于类鼻疽伯克霍尔德菌2号染色体上并编码σ因子RpoN同源物的rpoN2发生突变,并不会改变该细菌对氮和氨基酸的利用。然而,将类鼻疽伯克霍尔德菌的rpoN2导入缺乏rpoN的大肠杆菌菌株中,恢复了其利用氨基酸的能力。此外,对类鼻疽伯克霍尔德菌野生型及其rpoN2同基因突变体进行了比较性部分蛋白质组学分析,以阐明其氨基酸利用特性,这与其在互补试验中发现的功能相当。相比之下,rpoN2突变体在转录和翻译水平上katE的表达均降低。我们的研究结果表明,类鼻疽伯克霍尔德菌的RpoN2参与过氧化氢酶E表达调控的特定功能。