Identification of different promoters in the absA1–absA2 two-component system, a negative regulator of antibiotic production in Streptomyces coelicolor

Abstract

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.