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Genome-wide primary transcriptome landscape reveals the diversity of regulatory elements in archaeal genomes

ABSTRACT: We show the transcriptomic landscape of the H2-producing archaeon Thermococcus onnurineus NA1 in response to different growth conditions using primary and whole transcriptome sequencing. We identified 1,082 purine-rich transcription initiation sites along with well-conserved TATA boxes, A-rich B recognition elements (BREs), and promoter proximal element (PPE) motifs in promoter regions, a high pyrimidine nucleotide content (T/C) at the −1 position, and Shine-Dalgarno (SD) motifs (GGDGRD) in 5' untranslated regions (5' UTRs). In addition, along with differential transcript levels, 117 leaderless genes and 86 non-coding RNAs (ncRNAs) were identified, representing diverse cellular functions and potential regulatory functions during hydrogen production. Further comparative analysis of T. onnurineus upstream regulatory sequences with those of closely related archaeal genomes demonstrated that orthologous genes are expressed from highly conserved promoter sequences, however their upstream sequences for transcriptional and translational regulation are largely diverse. This study suggests that closely related archaea maintain orthologous gene functions, but may diversify the elements that regulate them to adapt to their unique environmental habitats.

ORGANISM(S): Thermococcus onnurineus

PROVIDER: GSE85760 | GEO | 2017/06/12

SECONDARY ACCESSION(S): PRJNA339284

REPOSITORIES: GEO

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Project description:Background: In prokaryotes, sigma factors are essential for the targeting of the transcription machinery to promoters. Various sigma factors have been described that recognize and bind to specific DNA sequence motifs in promoter sequences. The canonical sigma factor M-OM-^C70 is commonly involved in transcription of the cell's general housekeeping genes, which is mediated by the conserved M-OM-^C70 promoter sequence motifs. This study detects and predicts the general M-OM-^C70-promoter sequences in Lactobacillus plantarum WCFS1 using genome-wide analysis. The accuracy of the transcriptionally-active part of this promoter prediction was subsequently evaluated by correlating locations of predicted promoters with experimentally detected transcription starts sites (TSSs) using high-resolution tiling array transcriptome datasets. Results: To identify M-OM-^C70-related promoter sequences, we performed a genome-wide sequence motif scan of the L. plantarum WCFS1 genome focusing on the regions upstream of protein-encoding genes. We obtained several highly conserved motifs including those resembling the conserved M-OM-^C70-promoter consensus. Position weight matrices-based models of the recovered M-OM-^C70-promoter sequence motif were employed to identify 4746 motifs with significant similarity (p-value < 10-4) to the model-motif in the L. plantarum genome. Genome-wide transcription start site information deduced from whole genome tiling-array transcriptome datasets revealed 936 TSSs that were employed to validate the transcriptionally active fraction of these predicted promoters. In total, 578 predicted promoters were found in proximity (M-bM-^IM-$ 100 nucleotides) of the identified TSSs, showing a highly significant co-occurrence of predicted promoter and measured TSS (p-value < 10-23). An additional 224 predicted promoters was validated when the significant similarity to the model-motif was applied less strictly (10-4 M-bM-^IM-$ p-value < 10-3). Conclusions: High-resolution tiling arrays provide a suitable source for TSS detection at a genome-wide level, and allow experimental verification of in silico-predicted promoter sequence motifs. Triplicate hybridizations (technical replicates) of two L. plantarum WCFS1 samples (biological replicates) gathered from a fermentation run at OD600 of 1. Dye swap applied to one of the three technical replicates.

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Genome-wide primary transcriptome landscape reveals the diversity of regulatory elements in archaeal genomes

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