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- Title
Molecular motor tug-of-war regulates elongasome cell wall synthesis dynamics in Bacillus subtilis.
- Authors
Middlemiss, Stuart; Blandenet, Matthieu; Roberts, David M.; McMahon, Andrew; Grimshaw, James; Edwards, Joshua M.; Sun, Zikai; Whitley, Kevin D.; Blu, Thierry; Strahl, Henrik; Holden, Séamus
- Abstract
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. We found that the B. subtilis elongasome is highly processive and that processive synthesis events are frequently terminated by rapid reversal or extended pauses. We found that cellular levels of RodA regulate elongasome processivity, reversal and pausing. Our single-molecule data, together with stochastic simulations, show that elongasome dynamics and processivity are regulated by molecular motor tug-of-war competition between several, likely two, oppositely oriented peptidoglycan synthesis complexes associated with the MreB filament. Altogether these results demonstrate that molecular motor tug-of-war is a key regulator of elongasome dynamics in B. subtilis, which likely also regulates the cell shape via modulation of elongasome processivity. Most rod-shaped bacteria elongate using a protein complex, the elongasome, that inserts new cell wall material into the cell sidewall. Here, Middlemiss et al. track the movement of individual elongasomes around the circumference of Bacillus subtilis cells, providing evidence for a molecular motor tug-of-war competition between oppositely oriented cell-wall synthesis complexes.
- Subjects
MOLECULAR motor proteins; CARRIER proteins; CELL morphology; FLUORESCENCE microscopy; SURFACES (Technology)
- Publication
Nature Communications, 2024, Vol 15, Issue 1, p1
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-024-49785-x