Department of Microbiology, The Ohio State University, Columbus, Ohio, United States of America.
PLoS One. 2010 Jan 28;5(1):e8946. doi: 10.1371/journal.pone.0008946.
Escherichia and Salmonella encode SdiA, a transcription factor of the LuxR family that regulates genes in response to N-acyl homoserine lactones (AHLs) produced by other species of bacteria. E. coli genes that change expression in the presence of plasmid-encoded sdiA have been identified by several labs. However, many of these genes were identified by overexpressing sdiA on a plasmid and have not been tested for a response to sdiA produced from its natural position in the chromosome or for a response to AHL.
METHODOLOGY/PRINCIPAL FINDINGS: We determined that two important loci reported to respond to plasmid-based sdiA, ftsQAZ and acrAB, do not respond to sdiA expressed from its natural position in the chromosome or to AHLs. To identify genes that are regulated by chromosomal sdiA and/or AHLs, we screened 10,000 random transposon-based luciferase fusions in E. coli K-12 and a further 10,000 in E. coli O157:H7 for a response to AHL and then tested these genes for sdiA-dependence. We found that genes encoding the glutamate-dependent acid resistance system are up-regulated, and fliE is down-regulated, by sdiA. Gene regulation by sdiA of E. coli is only partially dependent upon AHL.
CONCLUSIONS/SIGNIFICANCE: The genes of E. coli that respond to plasmid-based expression of sdiA are largely different than those that respond to chromosomal sdiA and/or AHL. This has significant implications for determining the true function of AHL detection by E. coli.
大肠杆菌和沙门氏菌编码 SdiA,这是一种 LuxR 家族的转录因子,可响应其他细菌产生的 N-酰基高丝氨酸内酯 (AHL) 来调节基因表达。几个实验室已经确定了在质粒编码的 sdiA 存在下改变表达的大肠杆菌基因。然而,其中许多基因是通过在质粒上过度表达 sdiA 来鉴定的,尚未测试它们对天然染色体位置产生的 sdiA 的响应,也未测试它们对 AHL 的响应。
方法/主要发现:我们确定了两个被报道对基于质粒的 sdiA 有反应的重要基因座,ftsQAZ 和 acrAB,它们对天然染色体位置表达的 sdiA 或 AHL 没有反应。为了鉴定受染色体 sdiA 和/或 AHL 调节的基因,我们筛选了大肠杆菌 K-12 中的 10000 个随机转座子荧光素酶融合物,在大肠杆菌 O157:H7 中进一步筛选了 10000 个,以响应 AHL,然后测试了这些基因对 sdiA 的依赖性。我们发现,谷氨酸依赖的酸抗性系统的基因受 sdiA 上调,而 fliE 则受 sdiA 下调。大肠杆菌中 sdiA 的基因调控仅部分依赖于 AHL。
结论/意义:对质粒表达的 sdiA 有反应的大肠杆菌基因与对天然染色体 sdiA 和/或 AHL 有反应的基因有很大的不同。这对于确定大肠杆菌对 AHL 检测的真实功能具有重要意义。