Shaping Liposomes by Cell-Free Expressed Bacterial Microtubules

Johannes Kattan, Anne Doerr, Marileen Dogterom*, Christophe Danelon

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

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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 a 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.

Original languageEnglish
Number of pages9
JournalACS Synthetic Biology
Volume10
DOIs
Publication statusPublished - 2021

Keywords

  • artificial cell
  • cell-free gene expression
  • lipid vesicle
  • microtubule
  • morphogenesis
  • synthetic cell

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