
Shaping Liposomes by Cell-Free Expressed Bacterial Microtubules
Author(s) -
Johannes Kattan,
Anne Doerr,
Marileen Dogterom,
Christophe Danelon
Publication year - 2021
Publication title -
acs synthetic biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.156
H-Index - 66
ISSN - 2161-5063
DOI - 10.1021/acssynbio.1c00278
Subject(s) - microtubule , mreb , cytoskeleton , liposome , ftsz , vesicle , synthetic biology , gtp' , artificial cell , context (archaeology) , biology , microbiology and biotechnology , translation (biology) , dna , biophysics , membrane , chemistry , cell , biochemistry , computational biology , gene , messenger rna , paleontology , enzyme
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.