Phosphorylation-mediated regulation of the Bacillus anthracis phosphoglycerate mutase by the Ser/Thr protein kinase PrkC

Bacillus anthracis Ser/Thr protein kinase PrkC is necessary for phenotypic memory and spore germina-tion, and the loss of PrkC-dependent phosphorylation events affect the spore development. During sporulation, Bacillus sp. can store 3-Phosphoglycerate (3-PGA) that will be required at the onset of germination when ATP will be necessary. The Phosphoglycerate mutase (Pgm) catalyzes the isomeri-zation of 2-PGA and 3-PGA and is important for spore germination as a key metabolic enzyme that maintains 3-PGA pool at later events. Therefore, regulation of Pgm is important for an efficient spore germination process and metabolic switching. While the increased expression of Pgm in B. anthracis decreases spore germination efficiency, it remains unexplored if PrkC could directly influence Pgm ac-tivity. Here, we report the phosphorylation and regulation of Pgm by PrkC and its impact on Pgm stability and catalytic activity. Mass spectrometry revealed Pgm phosphorylation on seven threonine residues. In silico mutational analysis highlighted the role of Thr459 residue towards metal and substrate binding. Altogether, we demonstrated that PrkC-mediated Pgm phosphorylation negatively regulates its activity that is essential to maintain Pgm in its apo-like isoform before germination. This study advances the role of Pgm regulation that represents an important switch for B. anthracis resumption of metabolism and spore germination.(c) 2023 Elsevier Inc. All rights reserved.

Automated summary

PrkC protein kinase plays a vital role in the phenotypic memory and germination of Bacillus anthracis spores, and the loss of PrkC-dependent phosphorylation affects spore development. The key metabolic enzyme Phosphoglycerate mutase (Pgm) is involved in spore germination by catalyzing the isomerization of 2-PGA and 3-PGA. This study demonstrates that PrkC phosphorylates and regulates Pgm, negatively regulating its activity and maintaining its apo-like isoform before germination.