Department of Microbiology and Immunology, The University of Texas Health Science Center at San Antonio, Mail Code 7758, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900, USA.
Microb Pathog. 2011 Sep;51(3):133-41. doi: 10.1016/j.micpath.2011.04.009. Epub 2011 May 7.
The cyclic-AMP receptor protein (CRP) acts as a global regulatory protein among bacteria. Here, the CRP regulon has been defined in Aggregatibacter actinomycetemcomitans using microarray analysis of A. actinomycetemcomitans strain JP2 wild type cells compared to an isogenic crp deletion mutant. Genes whose expression levels changed at least 2-fold with p≤0.05 were considered significant. Of the 300 genes identified as being CRP-regulated, 139 were CRP-activated, including leukotoxin, with the remaining being CRP-repressed. The 300 genes represent 14.2% of ORFs probed which is significantly higher than what has been reported for CRP regulons in other bacteria. If the CRP-regulated genes are put into 17 functional classes, all 17 categories had at least 1 CRP-regulated gene. Several functional categories, mainly transport and binding proteins and energy metabolism proteins, were disproportionately represented in the CRP-regulated subset of genes relative to their overall representation in the genome. This is similar to the patterns seen in other bacteria. Finally, quantitative RT-PCR was used to show that the leukotoxin RNA levels were repressed 16-fold in the CRP mutant indicating that CRP activates leukotoxin transcription. However, this regulation appears to be acting through another regulatory protein since the leukotoxin promoter, unlike ∼129 other promoters of CRP-regulated genes, does not have a match to the consensus CRP-binding site. Several candidate genes for this intermediary transcription factor have been identified in the CRP regulon.
环腺苷酸受体蛋白 (CRP) 在细菌中充当全局调节蛋白。在这里,通过比较 Aggregatibacter actinomycetemcomitans 菌株 JP2 野生型细胞和同源 CRP 缺失突变体的微阵列分析,定义了 CRP 调控组。表达水平变化至少 2 倍且 p≤0.05 的基因被认为是显著的。在鉴定为 CRP 调控的 300 个基因中,有 139 个是 CRP 激活的,包括白细胞毒素,其余的是 CRP 抑制的。这 300 个基因代表了被探测到的 ORFs 的 14.2%,这明显高于其他细菌中 CRP 调控组的报告。如果将 CRP 调控基因分为 17 个功能类别,那么所有 17 个类别都至少有一个 CRP 调控基因。几个功能类别,主要是运输和结合蛋白和能量代谢蛋白,在 CRP 调控基因亚组中的代表性相对高于它们在基因组中的总体代表性。这与其他细菌中观察到的模式相似。最后,使用定量 RT-PCR 显示白细胞毒素 RNA 水平在 CRP 突变体中被抑制了 16 倍,表明 CRP 激活白细胞毒素转录。然而,这种调节似乎是通过另一种调节蛋白起作用的,因为白细胞毒素启动子与 CRP 调控基因的约 129 个其他启动子不同,没有与 CRP 结合位点的一致匹配。已经在 CRP 调控组中鉴定出了该中间转录因子的几个候选基因。