The Ser/Thr protein kinase PrkC imprints phenotypic memory in Bacillus anthracis spores by phosphorylating the glycolytic enzyme enolase

Abstract

Bacillus anthracis is the causative agent of anthrax in humans, bovine, and other animals. B. anthracis pathogenesis requires differentiation of dormant spores into vegetative cells. The spores inherit cellular components as phenotypic memory from the parent cell, and this memory plays a critical role in facilitating the spores' revival. Because metabolism initiates at the beginning of spore germination, here we metabolically reprogrammed B. anthracis cells to understand the role of glycolytic enzymes in this process. We show that increased expression of enolase (Eno) in the sporulating mother cell decreases germination efficiency. Eno is phosphorylated by the conserved Ser/Thr protein kinase PrkC which decreases the catalytic activity of Eno. We found that phosphorylation also regulates Eno expression and localization, thereby controlling the overall spore germination process. Using MS analysis, we identified the sites of phosphorylation in Eno, and substitution(s) of selected phosphorylation sites helped establish the functional correlation between phosphorylation and Eno activity. We propose that PrkC-mediated regulation of Eno may help sporulating B. anthracis cells in adapting to nutrient deprivation. In summary, to the best of our knowledge, our study provides the first evidence that in sporulating B. anthracis, PrkC imprints phenotypic memory that facilitates the germination process.

Integrative Analysis of Proteome and Transcriptome Dynamics during Bacillus subtilis Spore Revival

“…Thus, in-depth research on preexisting transcripts and proteins operative during spore revival is clearly important to understand long-term spore survival. Recently, enolase and alanine dehydrogenase present in dormant spores have been found to make up the spore’s phenotypic memory in Bacillus anthracis ( 10 ) and B. subtilis ( 9 ), respectively. These metabolic enzymes play a crucial role in optimally tuning the spore’s molecular composition to the metabolic needs that arise in efficiently progressing through spore germination and outgrowth.…”

“…To jump-start germination, the macromolecules needed are presynthesized during sporulation (8). The proteins that are transferred from the progenitor cell to the spore, the "tool-kit for life," are key to the spore's "memory" (9), which in turn modulates its germination response (10). Thus, in-depth research on preexisting transcripts and proteins operative during spore revival is clearly important to understand long-term spore survival.…”

Transcriptomics and Proteomics Analyses of the Responses of Propionibacterium acidipropionici to Metabolic and Evolutionary Manipulation

“…However, at the protein level, the expression of pfkA gene has not been identified but the enzyme aldo that converts 1.6-diphosphate-D-fructose to 3-phosphate-D-glyceraldehyde increased 2.24-fold. Another two key enzymes enolase and pyruvate phosphate dikinase catalyzing the formation of phosphoenolpyruvate from glycerate-2P in the glycolysis pathway was also observed to be significantly up-regulated ( Virmani et al, 2019 ). Generally, the absence of mRNA-protein correlation manifests that the relation between mRNA and protein is not strictly linear, but has a more inherent and complex dependence ( Wang et al, 2013 ).…”

Integrative analysis of proteome and transcriptome dynamics duringBacillus subtilisspore revival

“…The dormant spores harbour the transcriptional and translational machinery that might facilitates the early stages of outgrowth 5 . The protein cargo that transfers from progenitor cell to the spore, the ‘tool-kit for life’, is key to the spore’s memory 6 which in turn modulates its germination response 7 . Thus, clearly an in-depth research on pre-existing transcripts and proteins in spore revival is important to understand long-term stress survival.…”

“…Thus, clearly an in-depth research on pre-existing transcripts and proteins in spore revival is important to understand long-term stress survival. Recently, a few glycolytic enzymes and alanine dehydrogenase present in dormant spores have been identified to make up the spore’s phenotypic memory in Bacillus anthracis 7 .…”