RECTA: Regulon Identification Based on Comparative Genomics and Transcriptomics Analysis
Regulons, which serve as co-regulated gene groups contributing to the transcriptional regulation of microbial genomes, have the potential to aid in understanding of underlying regulatory mechanisms. In this study, we designed a novel computational pipeline, RECTA, for regulon prediction related to the gene regulatory network under certain conditions. To demonstrate the effectiveness of this tool, we implemented RECTA on Lactococcus lactis MG1363 data to elucidate acid-response regulons. Lactococcus lactis is one of the most important Gram-positive lactic acid-producing bacteria, widely used in food industry and has been proved to have advantages in oral delivery of drug and vaccine. The pipeline carries out differential gene expression, gene co-expression analysis, cis-regulatory motif finding, and comparative genomics to predict and validate regulons related to acid stress response. A total of 51 regulons were identified, 14 of which have computational-verified significance. Among these 14 regulons, five of them were computationally predicted to be connected with acid stress response with (i) known transcriptional factors in MEME suite database successfully mapped in Lactococcus lactis MG1363; and (ii) differentially expressed genes between pH values of 6.5 (control) and 5.1 (treatment). Validated by 36 literature confirmed acid stress response related proteins and genes, 33 genes in Lactococcus lactis MG1363 were found having orthologous genes using BLAST, associated to six regulons. An acid response related regulatory network was constructed, involving two trans-membrane proteins, eight regulons (llrA, llrC, hllA, ccpA, NHP6A, rcfB, regulons #8 and #39), nine functional modules, and 33 genes with orthologous genes known to be associated to acid stress. Our RECTA pipeline provides an effective way to construct a reliable gene regulatory network through regulon elucidation. The predicted response pathways could serve as promising candidates for better acid tolerance engineering in Lactococcus lactis. RECTA has strong application power and can be effectively applied to other bacterial genomes where the elucidation of the transcriptional regulation network is needed.
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