Clostridium thermocellum is a thermophilic bacterium recognized for its natural ability to effectively deconstruct cellulosic biomass. While there is a large body of studies on the genetic engineering of this bacterium and its physiology to-date, there is limited knowledge in the transcriptional regulation in this organism and thermophilic bacteria in general. The study herein is the first report of a high-throughput application of DNA-affinity purification sequencing (DAP-seq) to transcription factors (TFs) from a thermophile. We applied DAP-seq to >90 TFs in C. thermocellum and detected genome-wide binding sites for 11 of them. We then compiled and aligned DNA binding sequences from these TFs to deduce the primary DNA-binding sequence motifs for each TF. These binding motifs are further validated with electrophoretic mobility shift assay (EMSA) and are used to identify individual TFs’ regulatory targets in C. thermocellum. Our results led to the discovery of novel, uncharacterized TFs as well as homologues of previously studied TFs including RexA-, LexA- and LacI-type TFs. We then used these data to reconstruct gene regulatory networks for the 11 TFs individually, which resulted in a global network encompassing the TFs with some interconnections. As gene regulation governs and constrains how bacteria behave, our findings shed light on the roles of TFs delineated by their regulons, and potentially provides a means to enable rational, advanced genetic engineering of C. thermocellum and other organisms alike towards a desired phenotype.
2 Data Resources
|Cytoscape network file representing the predicted regulon for each TF and a global network encompassing all predicted regulons controlled by the TFs||275.39 KB||Document||Cytoscape network file representing the predicted regulon for each TF and a global network encompassing all predicted regulons controlled by the TFs|
|Supplementary File S1||157.81 KB||Document||Figures of reconstructed Clostridium thermocellum transcriptional regulatory networks showing the predicted regulon for each transcription factor (TF) and a global network encompassing all predicted regulons controlled by the TFs.|
• Apr • 19 2021
Cite This Dataset
Chou, Katherine, Skyler Hebdon, and Alida Gerritsen. 2021. "Genome-wide Transcription Factor DNA Binding Sites and Gene Regulatory Networks in Clostridium thermocellum." NREL Data Catalog. Golden, CO: National Renewable Energy Laboratory. Last updated: September 16, 2022. DOI: 10.7799/1778964.
About This Dataset
Center for Bioenergy Innovation (CBI)
Biomass & Biosciences Research (BBR)
Department of Energy (DOE)
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23). Biological Systems Science Division
Digital Object Identifier