Shaping Liposomes by Cell-Free Expressed Bacterial Microtubules
mediaposted on 29.09.2021, 15:37 by Johannes Kattan, Anne Doerr, Marileen Dogterom, Christophe Danelon
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.
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new building blockcontrolled shape changesde novo </work establishes bmtscompartmentalized inside liposomesbmts spontaneously interactb tubulins selfprotein filaments foundsynthetic cell biologysynthetic biologytubulins btubaprosthecobacter </synthetic cellshaping liposomesdeform liposomestranslation systemthus providingsynthesized btubareconstituted transcriptionpushing forcepotential routelipid membraneslipid compartmentlike environmentfunctionally expressedestablish polaritydna templatesdna segregationdivision machinerydisplay treadmillingdirectional transportdifferent morphologiescytoskeletal structuresbiophysical propertiesbacterial microtubulesactivated disassembly