Academic Journal
FtsW protein-protein interactions visualized in live Staphylococcus aureus cells by FLIM-FRET.
| Title: | FtsW protein-protein interactions visualized in live Staphylococcus aureus cells by FLIM-FRET. |
|---|---|
| Authors: | Meiresonne NY; Bacterial Cell Biology, Instituto de Tecnologia Química e Biológica António Xavier, Universidade NOVA de Lisboa, Oeiras, Portugal. nmeiresonne@itqb.unl.pt., Costa SF; Bacterial Cell Biology, Instituto de Tecnologia Química e Biológica António Xavier, Universidade NOVA de Lisboa, Oeiras, Portugal.; Single Molecule Microbiology, Instituto de Tecnologia Química e Biológica António Xavier, Universidade NOVA de Lisboa, Oeiras, Portugal., Schäper S; Bacterial Cell Biology, Instituto de Tecnologia Química e Biológica António Xavier, Universidade NOVA de Lisboa, Oeiras, Portugal., Ferreira MJ; Bacterial Cell Biology, Instituto de Tecnologia Química e Biológica António Xavier, Universidade NOVA de Lisboa, Oeiras, Portugal., Reed P; Bacterial Cell Biology, Instituto de Tecnologia Química e Biológica António Xavier, Universidade NOVA de Lisboa, Oeiras, Portugal., Hensel Z; Single Molecule Microbiology, Instituto de Tecnologia Química e Biológica António Xavier, Universidade NOVA de Lisboa, Oeiras, Portugal., Fernandes F; iBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.; i4HB-Institute for Health and Bioeconomy, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.; Bioengineering Department, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal., Pinho MG; Bacterial Cell Biology, Instituto de Tecnologia Química e Biológica António Xavier, Universidade NOVA de Lisboa, Oeiras, Portugal. mgpinho@itqb.unl.pt. |
| Source: | Nature communications [Nat Commun] 2026 May 09; Vol. 17 (1). Date of Electronic Publication: 2026 May 09. |
| Publication Type: | Journal Article |
| Language: | English |
| Journal Info: | Publisher: Nature Pub. Group Country of Publication: England NLM ID: 101528555 Publication Model: Electronic Cited Medium: Internet ISSN: 2041-1723 (Electronic) Linking ISSN: 20411723 NLM ISO Abbreviation: Nat Commun Subsets: MEDLINE |
| Imprint Name(s): | Original Publication: [London] : Nature Pub. Group |
| MeSH Terms: | Fluorescence Resonance Energy Transfer*/methods , Staphylococcus aureus*/metabolism , Bacterial Proteins*/metabolism , Bacterial Proteins*/genetics , Membrane Proteins*/metabolism , Membrane Proteins*/genetics , Cell Cycle Proteins*/metabolism, Peptidoglycan/biosynthesis ; Peptidoglycan/metabolism ; Microscopy, Fluorescence/methods ; Penicillin-Binding Proteins/metabolism ; Cell Wall/metabolism ; Protein Binding |
| Abstract: | The bacterial cell cycle relies on coordinated and dynamic interactions between division and peptidoglycan synthesis proteins. However, visualizing these interactions in vivo remains technically challenging. Here, we established fluorescence-lifetime imaging microscopy combined with Förster resonance energy transfer (FLIM-FRET) as a robust, spatially resolved technique to visualize protein interactions in living Staphylococcus aureus. We set up and validated the method using cytosolic and membrane-anchored control proteins, achieving FRET efficiencies of up to 40%. Using FLIM-FRET, we mapped protein interactions of the glycosyltransferase FtsW within the septal peptidoglycan-synthesizing complex. We confirmed its interaction with the cognate transpeptidase PBP1 and the regulatory protein DivIB. Notably, we found that FtsW also self-interacts, suggesting that septal peptidoglycan synthesis is performed by a complex of multimers, able to synthesize more than one glycan strand. Inhibition of peptidoglycan synthesis by directly targeting PBP1 with the beta-lactam antibiotic imipenem, but not by targeting the lipid II flippase, therefore depleting the FtsW-PBP1 substrate from the outer surface of the cell membrane, weakened FtsW-PBP1 interaction. This suggests that alterations in FtsW interactions result primarily from antibiotic-induced conformational changes or from uncoupling FtsW-PBP1 activities, resulting in the presence of uncrosslinked glycans, rather than merely from a loss of peptidoglycan synthesis activity. (© 2026. The Author(s).) |
| Competing Interests: | Competing interests: The authors declare no competing interests. |
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| Grant Information: | ERC-2022-ADG 101096393 EC | EU Framework Programme for Research and Innovation H2020 | H2020 Priority Excellent Science | H2020 European Research Council (H2020 Excellent Science - European Research Council); ERC-2017-CoG-771709 EC | EU Framework Programme for Research and Innovation H2020 | H2020 Priority Excellent Science | H2020 European Research Council (H2020 Excellent Science - European Research Council) |
| Substance Nomenclature: | 0 (Bacterial Proteins) 0 (Peptidoglycan) 0 (Membrane Proteins) 125724-13-2 (FtsW protein, Bacteria) 0 (Cell Cycle Proteins) 0 (Penicillin-Binding Proteins) |
| Entry Date(s): | Date Created: 20260509 Date Completed: 20260717 Latest Revision: 20260726 |
| Update Code: | 20260726 |
| PubMed Central ID: | PMC13376785 |
| DOI: | 10.1038/s41467-026-72752-7 |
| PMID: | 42106333 |
| Database: | MEDLINE |
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