PT  - JOURNAL ARTICLE
AU  - Johannes Kattan
AU  - Anne Doerr
AU  - Marileen Dogterom
AU  - Christophe Danelon
TI  - Shaping liposomes by cell-free expressed bacterial microtubules
AID  - 10.1101/2021.06.13.448053
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.06.13.448053
4099  - http://biorxiv.org/content/early/2021/06/13/2021.06.13.448053.short
4100  - http://biorxiv.org/content/early/2021/06/13/2021.06.13.448053.full
AB  - 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 StatementThe authors have declared no competing interest.