Reconstitution of a minimal machinery capable of assembling periplasmic type IV pili.

نویسندگان

  • Vivianne J Goosens
  • Andreas Busch
  • Michaella Georgiadou
  • Marta Castagnini
  • Katrina T Forest
  • Gabriel Waksman
  • Vladimir Pelicic
چکیده

Type IV pili (Tfp), which are key virulence factors in many bacterial pathogens, define a large group of multipurpose filamentous nanomachines widespread in Bacteria and Archaea. Tfp biogenesis is a complex multistep process, which relies on macromolecular assemblies composed of 15 conserved proteins in model gram-negative species. To improve our limited understanding of the molecular mechanisms of filament assembly, we have used a synthetic biology approach to reconstitute, in a nonnative heterologous host, a minimal machinery capable of building Tfp. Here we show that eight synthetic genes are sufficient to promote filament assembly and that the corresponding proteins form a macromolecular complex at the cytoplasmic membrane, which we have purified and characterized biochemically. Our results contribute to a better mechanistic understanding of the assembly of remarkable dynamic filaments nearly ubiquitous in prokaryotes.

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عنوان ژورنال:
  • Proceedings of the National Academy of Sciences of the United States of America

دوره 114 25  شماره 

صفحات  -

تاریخ انتشار 2017