Coupling Novel Probes with Molecular Localization Microscopy Reveals Cell Wall Homeostatic Mechanisms in Staphylococcus aureus
| Author(s) | Lund, Victoria A. | |
| Author(s) | Gangotra, Haneesh | |
| Author(s) | Zhao, Zhen | |
| Author(s) | Sutton, Joshua A. F. | |
| Author(s) | Wacnik, Katarzyna | |
| Author(s) | DeMeester, Kristen | |
| Author(s) | Liang, Hai | |
| Author(s) | Santiago, Cintia | |
| Author(s) | Grimes, Catherine Leimkuhler | |
| Author(s) | Jones, Simon | |
| Author(s) | Foster, Simon J. | |
| Date Accessioned | 2023-01-24T19:50:54Z | |
| Date Available | 2023-01-24T19:50:54Z | |
| Publication Date | 2022-11-22 | |
| Description | This article was originally published in ACS Chemical Biology. The version of record is available at: https://doi.org/10.1021/acschembio.2c00741 | |
| Abstract | Bacterial cell wall peptidoglycan is essential for viability, and its synthesis is targeted by antibiotics, including penicillin. To determine how peptidoglycan homeostasis controls cell architecture, growth, and division, we have developed novel labeling approaches. These are compatible with super-resolution fluorescence microscopy to examine peptidoglycan synthesis, hydrolysis, and the localization of the enzymes required for its biosynthesis (penicillin binding proteins (PBPs)). Synthesis of a cephalosporin-based fluorescent probe revealed a pattern of PBPs at the septum during division, supporting a model of dispersed peptidoglycan synthesis. Metabolic and hydroxylamine-based probes respectively enabled the synthesis of glycan strands and associated reducing termini of the peptidoglycan to be mapped. Foci and arcs of reducing termini appear as a result of both synthesis of glycan strands and glucosaminidase activity of the major peptidoglycan hydrolase, SagB. Our studies provide molecular level details of how essential peptidoglycan dynamics are controlled during growth and division. | |
| Sponsor | This research was funded in whole, or in part, by the Wellcome Trust (212197/Z/19/Z). For the purpose of open access, the authors have applied a CC BY public copyright licence to any Author Accepted Manuscript version arising from this submission. This work was also funded by the Medical Research Council (MR/N002679/1), UKRI Strategic Priorities Fund (EP/T002778/1), and the Wellcome Trust (212197/Z/19/Z). C.L.G. thanks the NIH Glycoscience Commonfund (NIH U01CA221230), and K.E.D. was supported by an NIH T32 Training Grant (T32GM133395). We are grateful to M. Mohaghegh for the provision of strains. The graphical abstract was created using BioRender.com. | |
| Citation | Lund, Victoria A., Haneesh Gangotra, Zhen Zhao, Joshua A. F. Sutton, Katarzyna Wacnik, Kristen DeMeester, Hai Liang, et al. “Coupling Novel Probes with Molecular Localization Microscopy Reveals Cell Wall Homeostatic Mechanisms in Staphylococcus Aureus.” ACS Chemical Biology 17, no. 12 (December 16, 2022): 3298–3305. https://doi.org/10.1021/acschembio.2c00741. | |
| ISSN | 1554-8937 | |
| URL | https://udspace.udel.edu/handle/19716/32144 | |
| Language | en_US | |
| Publisher | ACS Chemical Biology | |
| Keywords | carbohydrates | |
| Keywords | cells | |
| Keywords | chemical biology | |
| Keywords | labeling | |
| Keywords | peptides and proteins | |
| Title | Coupling Novel Probes with Molecular Localization Microscopy Reveals Cell Wall Homeostatic Mechanisms in Staphylococcus aureus | |
| Type | Article |
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