ArticleNucleic acids research2025
Characterization of the MgsR-dependent promoter structure in Bacillus subtilis-application of a novel pHIS plasmid-based screening system for promoter element analysis.
Article in Nucleic acids research, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.
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Who cites it
2 citing papers in PubMed.
- Optimization Strategy of Expression Vectors and Regulatory Elements for Enhanced Protein Production inInternational journal of molecular sciences · 2025Review
- Requirement of ClpX for CtsR dissociation from its operator elements upon heat stress inFrontiers in microbiology · 2025Article
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Authors and funding
8 authors.
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Abstract
Transcriptional control is a finely tuned process, crucial for adaptation, differentiation, and survival. However, deciphering the contribution of promoter elements to overall promoter strength is tedious. We present the novel plasmid-based screening system pHIS (pHigh Screen) as a tool for easy and quantitative monitoring of promoter activity via fluorescence reporter gene expression for a reliable in vivo promoter analysis in real time. As a proof of principle, we first validated the functionality of our pHIS system by analyzing the known Spx operator of a SigA-type promoter in Bacillus subtilis. Then, the pHIS plasmid was used to characterize a new operator element, which was predicted by in silico analysis of SigB-type promoters controlled by the Spx paralogue MgsR. We found evidence for (i) an intrinsic promoter silencer element located upstream of the SigB-type -35 core region, preventing target gene expression in the absence of MgsR and (ii) an MgsR-dependent antagonist enhancer element necessary to overcome the negative element and to achieve full stress-dependent promoter activation. Our data demonstrate the universal applicability of our novel reporter gene system pHIS and support an RNA polymerase repositioning model for transcription initiation at MgsR-regulated SigB-type promoters in B. subtilis.
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