Rodionov Dmitry A, Gelfand Mikhail S, Hugouvieux-Cotte-Pattat Nicole
State Scientific Centre GosNIIGenetika, Moscow, 117545, Russia.
Institute for Problems of Information Transmission, Russian Academy of Sciences, Bolshoy Karetny per. 19, Moscow GSP-4, 127994, Russia.
Microbiology (Reading). 2004 Nov;150(Pt 11):3571-3590. doi: 10.1099/mic.0.27041-0.
In the plant-pathogenic enterobacterium Erwinia chrysanthemi, almost all known genes involved in pectin catabolism are controlled by the transcriptional regulator KdgR. In this study, the comparative genomics approach was used to analyse the KdgR regulon in completely sequenced genomes of eight enterobacteria, including Erw. chrysanthemi, and two Vibrio species. Application of a signal recognition procedure complemented by operon structure and protein sequence analysis allowed identification of new candidate genes of the KdgR regulon. Most of these genes were found to be controlled by the cAMP-receptor protein, a global regulator of catabolic genes. At the next step, regulation of these genes in Erw. chrysanthemi was experimentally verified using in vivo transcriptional fusions and an attempt was made to clarify the functional role of the predicted genes in pectin catabolism. Interestingly, it was found that the KdgR protein, previously known as a repressor, positively regulates expression of two new members of the regulon, phosphoenolpyruvate synthase gene ppsA and an adjacent gene, ydiA, of unknown function. Other predicted regulon members, namely chmX, dhfX, gntB, pykF, spiX, sotA, tpfX, yeeO and yjgK, were found to be subject to classical negative regulation by KdgR. Possible roles of newly identified members of the Erw. chrysanthemi KdgR regulon, chmX, dhfX, gntDBMNAC, spiX, tpfX, ydiA, yeeO, ygjV and yjgK, in pectin catabolism are discussed. Finally, complete reconstruction of the KdgR regulons in various gamma-proteobacteria yielded a metabolic map reflecting a globally conserved pathway for the catabolism of pectin and its derivatives with variability in transport and enzymic capabilities among species. In particular, possible non-orthologous substitutes of isomerase KduI and a new oligogalacturonide transporter in the Vibrio species were detected.
在植物致病性肠道细菌菊欧文氏菌(Erwinia chrysanthemi)中,几乎所有已知参与果胶分解代谢的基因都受转录调节因子KdgR的控制。在本研究中,采用比较基因组学方法分析了包括菊欧文氏菌在内的8种肠道细菌以及2种弧菌的全序列基因组中的KdgR调控子。通过操纵子结构和蛋白质序列分析辅助的信号识别程序,得以鉴定出KdgR调控子的新候选基因。发现这些基因中的大多数受分解代谢基因的全局调节因子——环腺苷酸受体蛋白的控制。下一步,利用体内转录融合实验验证了菊欧文氏菌中这些基因的调控情况,并尝试阐明预测基因在果胶分解代谢中的功能作用。有趣的是,发现先前被认为是阻遏物的KdgR蛋白正向调节调控子的两个新成员——磷酸烯醇式丙酮酸合酶基因ppsA和一个功能未知的相邻基因ydiA的表达。其他预测的调控子成员,即chmX、dhfX、gntB、pykF、spiX、sotA、tpfX、yeeO和yjgK,被发现受KdgR的经典负调控。讨论了菊欧文氏菌KdgR调控子新鉴定成员chmX、dhfX、gntDBMNAC、spiX、tpfX、ydiA、yeeO、yjgV和yjgK在果胶分解代谢中的可能作用。最后,对各种γ-变形菌中的KdgR调控子进行完整重建,得到了一个代谢图谱,该图谱反映了果胶及其衍生物分解代谢的全局保守途径,不同物种在转运和酶促能力方面存在差异。特别是,检测到弧菌中异构酶KduI的可能非直系同源替代物和一种新的寡聚半乳糖醛酸转运蛋白。