3 years ago

Reconstitution of a minimal machinery capable of assembling periplasmic type IV pili [Microbiology]

Reconstitution of a minimal machinery capable of assembling periplasmic type IV pili [Microbiology]
Vladimir Pelicic, Marta Castagnini, Gabriel Waksman, Vivianne J. Goosens, Katrina T. Forest, Michaella Georgiadou, Andreas Busch

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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