Differential GTP-dependent in-vitro polymerization of recombinant Physcomitrella FtsZ proteins
Abstract Bacterial cell division and plant chloroplast division require selfassembling Filamentous temperature-sensitive Z (FtsZ) proteins. FtsZ proteins are GTPases sharing structural and biochemical similarities with eukaryotic tubulin. In the moss Physcomitrella, the morphology of the FtsZ polyme...
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Nature Portfolio
2025-01-01
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| Online Access: | https://doi.org/10.1038/s41598-024-85077-6 |
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| author | Stella W. L. Milferstaedt Marie Joest Lennard L. Bohlender Sebastian N. W. Hoernstein Buğra Özdemir Eva L. Decker Chris van der Does Ralf Reski |
| author_facet | Stella W. L. Milferstaedt Marie Joest Lennard L. Bohlender Sebastian N. W. Hoernstein Buğra Özdemir Eva L. Decker Chris van der Does Ralf Reski |
| author_sort | Stella W. L. Milferstaedt |
| collection | DOAJ |
| description | Abstract Bacterial cell division and plant chloroplast division require selfassembling Filamentous temperature-sensitive Z (FtsZ) proteins. FtsZ proteins are GTPases sharing structural and biochemical similarities with eukaryotic tubulin. In the moss Physcomitrella, the morphology of the FtsZ polymer networks varies between the different FtsZ isoforms. The underlying mechanism and foundation of the distinct networks is unknown. Here, we investigated the interaction of Physcomitrella FtsZ2-1 with FtsZ1 isoforms via co-immunoprecipitation and mass spectrometry, and found protein-protein interaction in vivo. We tagged FtsZ1-2 and FtsZ2-1 with different fluorophores and expressed both in E. coli, which led to the formation of defined structures within the cells and to an influence on bacterial cell division and morphology. Furthermore, we have optimized the purification protocols for FtsZ1-2 and FtsZ2-1 expressed in E. coli and characterized their GTPase activity and polymerization in vitro. Both FtsZ isoforms showed GTPase activity. Stoichiometric mixing of both proteins led to a significantly increased GTPase activity, indicating a synergistic interaction between them. In light scattering assays, we observed GTP-dependent assembly of FtsZ1-2 and of FtsZ2-1 in a protein concentration dependent manner. Stoichiometric mixing of both proteins resulted in significantly faster polymerization, again indicating a synergistic interaction between them. Under the same conditions used for GTPase and light scattering assays both FtsZ isoforms formed filaments in a GTP-dependent manner as visualized by transmission electron microscopy (TEM). Taken together, our results reveal that Physcomitrella FtsZ1-2 and FtsZ2-1 are functionally different, can synergistically interact in vivo and in vitro, and differ in their properties from FtsZ proteins from bacteria, archaea and vascular plants. |
| format | Article |
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| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
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| series | Scientific Reports |
| spelling | doaj-art-9ba6ecf68a31455faecf162270d57a2e2025-01-26T12:27:13ZengNature PortfolioScientific Reports2045-23222025-01-0115111610.1038/s41598-024-85077-6Differential GTP-dependent in-vitro polymerization of recombinant Physcomitrella FtsZ proteinsStella W. L. Milferstaedt0Marie Joest1Lennard L. Bohlender2Sebastian N. W. Hoernstein3Buğra Özdemir4Eva L. Decker5Chris van der Does6Ralf Reski7Plant Biotechnology, Faculty of Biology, University of FreiburgMolecular Biology of Archaea, Faculty of Biology, University of FreiburgPlant Biotechnology, Faculty of Biology, University of FreiburgPlant Biotechnology, Faculty of Biology, University of FreiburgPlant Biotechnology, Faculty of Biology, University of FreiburgPlant Biotechnology, Faculty of Biology, University of FreiburgMolecular Biology of Archaea, Faculty of Biology, University of FreiburgPlant Biotechnology, Faculty of Biology, University of FreiburgAbstract Bacterial cell division and plant chloroplast division require selfassembling Filamentous temperature-sensitive Z (FtsZ) proteins. FtsZ proteins are GTPases sharing structural and biochemical similarities with eukaryotic tubulin. In the moss Physcomitrella, the morphology of the FtsZ polymer networks varies between the different FtsZ isoforms. The underlying mechanism and foundation of the distinct networks is unknown. Here, we investigated the interaction of Physcomitrella FtsZ2-1 with FtsZ1 isoforms via co-immunoprecipitation and mass spectrometry, and found protein-protein interaction in vivo. We tagged FtsZ1-2 and FtsZ2-1 with different fluorophores and expressed both in E. coli, which led to the formation of defined structures within the cells and to an influence on bacterial cell division and morphology. Furthermore, we have optimized the purification protocols for FtsZ1-2 and FtsZ2-1 expressed in E. coli and characterized their GTPase activity and polymerization in vitro. Both FtsZ isoforms showed GTPase activity. Stoichiometric mixing of both proteins led to a significantly increased GTPase activity, indicating a synergistic interaction between them. In light scattering assays, we observed GTP-dependent assembly of FtsZ1-2 and of FtsZ2-1 in a protein concentration dependent manner. Stoichiometric mixing of both proteins resulted in significantly faster polymerization, again indicating a synergistic interaction between them. Under the same conditions used for GTPase and light scattering assays both FtsZ isoforms formed filaments in a GTP-dependent manner as visualized by transmission electron microscopy (TEM). Taken together, our results reveal that Physcomitrella FtsZ1-2 and FtsZ2-1 are functionally different, can synergistically interact in vivo and in vitro, and differ in their properties from FtsZ proteins from bacteria, archaea and vascular plants.https://doi.org/10.1038/s41598-024-85077-6CytoskeletonFtsZ polymerizationGTPasePhyscomitriumPlastoskeletonSelf-assembly |
| spellingShingle | Stella W. L. Milferstaedt Marie Joest Lennard L. Bohlender Sebastian N. W. Hoernstein Buğra Özdemir Eva L. Decker Chris van der Does Ralf Reski Differential GTP-dependent in-vitro polymerization of recombinant Physcomitrella FtsZ proteins Scientific Reports Cytoskeleton FtsZ polymerization GTPase Physcomitrium Plastoskeleton Self-assembly |
| title | Differential GTP-dependent in-vitro polymerization of recombinant Physcomitrella FtsZ proteins |
| title_full | Differential GTP-dependent in-vitro polymerization of recombinant Physcomitrella FtsZ proteins |
| title_fullStr | Differential GTP-dependent in-vitro polymerization of recombinant Physcomitrella FtsZ proteins |
| title_full_unstemmed | Differential GTP-dependent in-vitro polymerization of recombinant Physcomitrella FtsZ proteins |
| title_short | Differential GTP-dependent in-vitro polymerization of recombinant Physcomitrella FtsZ proteins |
| title_sort | differential gtp dependent in vitro polymerization of recombinant physcomitrella ftsz proteins |
| topic | Cytoskeleton FtsZ polymerization GTPase Physcomitrium Plastoskeleton Self-assembly |
| url | https://doi.org/10.1038/s41598-024-85077-6 |
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