Article: article from journal or magazin.
Heterochronic phosphorelay gene expression as a source of heterogeneity in Bacillus subtilis spore formation.
Journal of Bacteriology
In response to limiting nutrient sources and cell density signals, Bacillus subtilis can differentiate and form highly resistant endospores. Initiation of spore development is governed by the master regulator Spo0A, which is activated by phosphorylation via a multicomponent phosphorelay. Interestingly, only part of a clonal population will enter this developmental pathway, a phenomenon known as sporulation bistability or sporulation heterogeneity. How sporulation heterogeneity is established is largely unknown. To investigate the origins of sporulation heterogeneity, we constructed promoter-green fluorescent protein (GFP) fusions to the main phosphorelay genes and perturbed their expression levels. Using time-lapse fluorescence microscopy and flow cytometry, we showed that expression of the phosphorelay genes is distributed in a unimodal manner. However, single-cell trajectories revealed that phosphorelay gene expression is highly dynamic or "heterochronic" between individual cells and that stochasticity of phosphorelay gene transcription might be an important regulatory mechanism for sporulation heterogeneity. Furthermore, we showed that artificial induction or depletion of the phosphorelay phosphate flow results in loss of sporulation heterogeneity. Our data suggest that sporulation heterogeneity originates from highly dynamic and variable gene activity of the phosphorelay components, resulting in large cell-to-cell variability with regard to phosphate input into the system. These transcriptional and posttranslational differences in phosphorelay activity appear to be sufficient to generate a heterogeneous sporulation signal without the need of the positive-feedback loop established by the sigma factor SigH.
Bacillus subtilis/genetics, Bacillus subtilis/growth & development, Bacterial Proteins/genetics, Bacterial Proteins/metabolism, Flow Cytometry, Gene Expression Regulation, Bacterial/genetics, Gene Expression Regulation, Bacterial/physiology, Microscopy, Fluorescence, Models, Biological, Phosphoprotein Phosphatases/genetics, Phosphoprotein Phosphatases/metabolism, Protein Kinases/genetics, Protein Kinases/metabolism, Spores, Bacterial/genetics, Spores, Bacterial/growth & development, Transcription Factors/genetics, Transcription Factors/metabolism
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