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Shaping liposomes by cell-free expressed bacterial microtubules

Johannes Kattan, Anne Doerr, Marileen Dogterom, Christophe Danelon
doi: https://doi.org/10.1101/2021.06.13.448053
Johannes Kattan
Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, van der Maasweg 9, 2629 HZ, Delft, The Netherlands
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Anne Doerr
Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, van der Maasweg 9, 2629 HZ, Delft, The Netherlands
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Marileen Dogterom
Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, van der Maasweg 9, 2629 HZ, Delft, The Netherlands
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  • For correspondence: M.Dogterom@tudelft.nl C.J.A.Danelon@tudelft.nl
Christophe Danelon
Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, van der Maasweg 9, 2629 HZ, Delft, The Netherlands
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  • For correspondence: M.Dogterom@tudelft.nl C.J.A.Danelon@tudelft.nl
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ABSTRACT

Genetic control over a cytoskeletal network inside lipid vesicles offers a potential route to controlled shape changes and DNA segregation in synthetic cell biology. Bacterial microtubules (bMTs) are protein filaments found in bacteria of the genus Prosthecobacter. They are formed by the tubulins BtubA and BtubB which polymerize in the presence of GTP. Here, we show that the tubulins BtubA/B can be functionally expressed from DNA templates in a reconstituted transcription-translation system, thus providing a cytosol-like environment to study their biochemical and biophysical properties. We found that bMTs spontaneously interact with lipid membranes and display treadmilling. When compartmentalized inside liposomes, de novo synthesized BtubA/B tubulins self-organize into cytoskeletal structures of different morphologies. Moreover, bMTs can exert a pushing force on the membrane and deform liposomes, a phenomenon that can be reversed by light-activated disassembly of the filaments. Our work establishes bMTs as a new building block in synthetic biology. In the context of creating a synthetic cell, bMTs could help shape the lipid compartment, establish polarity or directional transport, and assist the division machinery.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted June 13, 2021.
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Shaping liposomes by cell-free expressed bacterial microtubules
Johannes Kattan, Anne Doerr, Marileen Dogterom, Christophe Danelon
bioRxiv 2021.06.13.448053; doi: https://doi.org/10.1101/2021.06.13.448053
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Shaping liposomes by cell-free expressed bacterial microtubules
Johannes Kattan, Anne Doerr, Marileen Dogterom, Christophe Danelon
bioRxiv 2021.06.13.448053; doi: https://doi.org/10.1101/2021.06.13.448053

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