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C-terminal eYFP fusion impairs Escherichia coli MinE function

Palanisamy, Navaneethan
; 
Öztürk, Mehmet Ali
; 
Akmeriç, Emir Bora
; 
Di Ventura, Barbara
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Affiliation
University of Freiburg; University of Heidelberg
EPub Date
Publication Date
2020-01-10
Submitted Date
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Chester Medical School
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Abstract
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 <i>in vivo</i>, <i>in vitro</i> and <i>in silico</i> 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. <i>In silico</i> 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.
Citation
Palanisamy, N., Öztürk, M. A., Akmeriç, E. B., & Di Ventura, B. (2020). C-terminal eYFP fusion impairs Escherichia coli MinE function. Open Biology, 10(5), 200010. https://doi.org/10.1098/rsob.200010
Publisher
The Royal Society
Journal
Open Biology
Research Unit
URI
http://hdl.handle.net/10034/629474
DOI
10.1098/rsob.200010
PubMed ID
PubMed Central ID
Type
Article
Language
en
Description
Series/Report no.
ISSN
2046-2441
EISSN
ISBN
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Sponsors
Deutsche Forschungsgemeinschaft (DFG); Bundesministerium für Bildung und Forschung (BMBF); German Federal and State Governments (BIOSS)
Additional Links
https://royalsocietypublishing.org/doi/10.1098/rsob.200010