Instituto de Biotecnología de León, INBIOTEC, Parque Científico de León, Av. Real, 1, 24006 León, Spain.
Mol Genet Genomics. 2013 Feb;288(1-2):39-48. doi: 10.1007/s00438-012-0728-2. Epub 2012 Dec 18.
The absA1-absA2 genes encode a two-component system that negatively regulates the transcription of multiple antibiotic gene clusters of Streptomyces coelicolor. The microarray dataset time series of a S. coelicolor M145 bioreactor culture indicated that the transcription values of absA2 were approximately four times higher than those of absA1 throughout the time course of the culture. The co-transcription of absA1 and absA2 genes has been previously shown, although an independent absA2 promoter was not detected. In this study, we show by different technical approaches that the absA1-absA2 operon is transcribed from at least two promoters, the first producing a read-through transcript that includes both absA1 and absA2 genes and the second including only the absA2 gene. An absA2 mRNA 5' end was mapped by primer extension and confirmed as TSS by deep sequencing in combination with TEX. Promoter-probe analyses detected promoter activity in both the absA1 and absA2 upstream regions. The absA2 upstream region showed a higher promoter activity, at least sevenfold higher than that of absA1. Furthermore, the absA2 gene may contain at least two additional promoters as shown by deep sequencing analyses. All together this work contributes to the understanding of the complex transcriptional regulation of these antibiotic regulators genes in S. coelicolor.
absA1-absA2 基因编码一个双组分系统,该系统负调控链霉菌协同素中多个抗生素基因簇的转录。链霉菌协同素 M145 生物反应器培养的微阵列数据集时间序列表明,在整个培养过程中,absA2 的转录值大约是 absA1 的四倍。虽然没有检测到独立的 absA2 启动子,但先前已经显示了 absA1 和 absA2 基因的共转录。在这项研究中,我们通过不同的技术方法表明,absA1-absA2 操纵子至少由两个启动子转录,第一个启动子产生一个通读转录本,其中包含 absA1 和 absA2 基因,第二个启动子只包含 absA2 基因。引物延伸和结合 TEX 的深度测序将 absA2 mRNA 5' 端映射,并通过确认其为 TSS。启动子探针分析检测到 absA1 和 absA2 上游区域的启动子活性。absA2 上游区域显示出更高的启动子活性,至少比 absA1 高七倍。此外,如深度测序分析所示,absA2 基因可能包含至少两个额外的启动子。总的来说,这项工作有助于理解链霉菌协同素中这些抗生素调节剂基因的复杂转录调控。