C-terminal eYFP fusion impairs Escherichia coli MinE function
The Escherichia coli Min system plays an important role in the proper placement of the septum ring at mid-cell during cell division. MinE forms a pole-to-pole spatial oscillator with the membrane-bound ATPase MinD, resulting in MinD concentration being the lowest at mid-cell. MinC, the direct inhibi...
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| Main Authors: | , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
27 May 2020
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| In: |
Open biology
Year: 2020, Volume: 10, Issue: 5, Pages: 1-14 |
| ISSN: | 2046-2441 |
| DOI: | 10.1098/rsob.200010 |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1098/rsob.200010 Verlag, lizenzpflichtig, Volltext: https://royalsocietypublishing.org/doi/10.1098/rsob.200010 
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| Author Notes: | Navaneethan Palanisamy, Mehmet Ali Öztürk, Emir Bora Akmeriç and Barbara Di Ventura |
| Summary: | The Escherichia coli Min system plays an important role in the proper placement of the septum ring at mid-cell during cell division. MinE forms a pole-to-pole spatial oscillator with the membrane-bound ATPase MinD, resulting in MinD concentration being the lowest at mid-cell. MinC, the direct inhibitor of the septum initiator protein FtsZ, forms a complex with MinD at the membrane, mirroring its polar gradients. Therefore, MinC-mediated FtsZ inhibition occurs away from mid-cell. Min oscillations are often studied in living cells by time-lapse microscopy using fluorescently labelled Min proteins. Here, we show that, despite permitting oscillations to occur in a range of protein concentrations, the enhanced yellow fluorescent protein (eYFP) C-terminally fused to MinE impairs its function. Combining in vivo, in vitro and in silico approaches, we demonstrate that eYFP compromises the ability of MinE to displace MinC from MinD, to stimulate MinD ATPase activity and to directly bind to the membrane. Moreover, we reveal that MinE-eYFP is prone to aggregation. In silico analyses predict that other fluorescent proteins are also likely to compromise several functionalities of MinE, suggesting that the results presented here are not specific to eYFP. |
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| Item Description: | Gesehen am 19.03.2021 |
| Physical Description: | Online Resource |
| ISSN: | 2046-2441 |
| DOI: | 10.1098/rsob.200010 |