da Silva Neto José F, Koide Tie, Gomes Suely L, Marques Marilis V
Departmento de Microbiologia, Universidade de São Paulo, Brazil.
J Bacteriol. 2007 Jan;189(2):551-60. doi: 10.1128/JB.00986-06. Epub 2006 Nov 10.
Genome sequence analysis of the bacterium Xylella fastidiosa revealed the presence of two genes, named rpoE and rseA, predicted to encode an extracytoplasmic function (ECF) sigma factor and an anti-sigma factor, respectively. In this work, an rpoE null mutant was constructed in the citrus strain J1a12 and shown to be sensitive to exposure to heat shock and ethanol. To identify the X. fastidiosa sigma(E) regulon, global gene expression profiles were obtained by DNA microarray analysis of bacterial cells under heat shock, identifying 21 sigma(E)-dependent genes. These genes encode proteins belonging to different functional categories, such as enzymes involved in protein folding and degradation, signal transduction, and DNA restriction modification and hypothetical proteins. Several putative sigma(E)-dependent promoters were mapped by primer extension, and alignment of the mapped promoters revealed a consensus sequence similar to those of ECF sigma factor promoters of other bacteria. Like other ECF sigma factors, rpoE and rseA were shown to comprise an operon in X. fastidiosa, together with a third open reading frame (XF2241). However, upon heat shock, rpoE expression was not induced, while rseA and XF2241 were highly induced at a newly identified sigma(E)-dependent promoter internal to the operon. Therefore, unlike many other ECF sigma factors, rpoE is not autoregulated but instead positively regulates the gene encoding its putative anti-sigma factor.
对木质部难养菌(Xylella fastidiosa)的基因组序列分析显示,存在两个基因,分别命名为rpoE和rseA,预计它们分别编码一种胞质外功能(ECF)σ因子和一种抗σ因子。在这项研究中,在柑橘菌株J1a12中构建了rpoE基因缺失突变体,并发现其对热休克和乙醇暴露敏感。为了鉴定木质部难养菌的σ(E)调控子,通过对热休克条件下的细菌细胞进行DNA微阵列分析获得了全局基因表达谱,鉴定出21个依赖σ(E)的基因。这些基因编码属于不同功能类别的蛋白质,如参与蛋白质折叠和降解、信号转导、DNA限制修饰的酶以及假定蛋白。通过引物延伸定位了几个假定的依赖σ(E)的启动子,对定位的启动子进行比对发现了一个与其他细菌的ECF σ因子启动子相似的共有序列。与其他ECF σ因子一样,rpoE和rseA在木质部难养菌中与第三个开放阅读框(XF2241)一起构成一个操纵子。然而,在热休克时,rpoE的表达未被诱导,而rseA和XF2241在操纵子内部新鉴定的依赖σ(E)的启动子处被高度诱导。因此,与许多其他ECF σ因子不同,rpoE不是自我调节的,而是正向调节编码其假定抗σ因子的基因。