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

Goosens, V.J. and Busch, Andreas and Georgiadou, M. and Castagnini, M. and Forest, K.T. and Waksman, Gabriel and Pelicic, V. (2017) Reconstitution of a minimal machinery capable of assembling periplasmic type IV pili. Proceedings of the National Academy of Sciences of the United States of America 114 (25), E4978-E4986. ISSN 0027-8424.

Abstract

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 heter- ologous 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 con- tribute to a better mechanistic understanding of the assembly of remarkable dynamic filaments nearly ubiquitous in prokaryotes.

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