biostudies-literatureMiddlemiss SRCUK | Biotechnology and Biological Sciences Research CouncilWellcome TrustBiotechnology and Biological Sciences Research Council5411https://www.ebi.ac.uk/biostudies/studies/S-EPMC11208587biostudies-literatureUnknown15(1)Most rod-shaped bacteria elongate by inserting new cell wall material into the inner surface of the cell sidewall. This is performed by class A penicillin binding proteins (PBPs) and a highly conserved protein complex, the elongasome, which moves processively around the cell circumference and inserts long glycan strands that act as barrel-hoop-like reinforcing structures, thereby giving rise to a rod-shaped cell. However, it remains unclear how elongasome synthesis dynamics and termination events are regulated to determine the length of these critical cell-reinforcing structures. To address this, we developed a method to track individual elongasome complexes around the entire circumference of Bacillus subtilis cells for minutes-long periods using single-molecule fluorescence microscopy. WeNature communicationsMolecular motor tug-of-war regulates elongasome cell wall synthesis dynamics in Bacillus subtilis.PMC11208587BB/X001482/1BB/X001512/1BB/M011186/1206670/Z/17/ZRoberts DMMcMahon ABlandenet MEdwards JMWhitley KDStrahl HGrimshaw JBlu TMiddlemiss SSun ZHolden SfalseMolecular motor tug-of-war regulates elongasome cell wall synthesis dynamics in Bacillus subtilis.Most rod-shaped bacteria elongate by inserting new cell wall material into the inner surface of the cell sidewall. This is performed by class A penicillin binding proteins (PBPs) and a highly conserved protein complex, the elongasome, which moves processively around the cell circumference and inserts long glycan strands that act as barrel-hoop-like reinforcing structures, thereby giving rise to a rod-shaped cell. However, it remains unclear how elongasome synthesis dynamics and termination events are regulated to determine the length of these critical cell-reinforcing structures. To address this, we developed a method to track individual elongasome complexes around the entire circumference of Bacillus subtilis cells for minutes-long periods using single-molecule fluorescence microscopy. We2024-01-01T00:00:00Z2024 Jun2025-04-04T21:22:31.628Z2025-04-04T21:22:31.628ZS-EPMC112085873892633610.1038/s41467-024-49785-x