Multiple conformations facilitate PilT function in the type IV pilus.

McCallum, Matthew; Benlekbir, Samir; Nguyen, Sheryl; Tammam, Stephanie; Rubinstein, John L; Burrows, Lori L; Howell, P Lynne · Nat Commun · 2019

basic_science · Level V

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Abstract

Type IV pilus-like systems are protein complexes that polymerize pilin fibres. They are critical for virulence in many bacterial pathogens. Pilin polymerization and depolymerization are powered by motor ATPases of the PilT/VirB11-like family. This family is thought to operate with C<sub>2</sub> symmetry; however, most of these ATPases crystallize with either C<sub>3</sub> or C<sub>6</sub> symmetric conformations. The relevance of these conformations is unclear. Here, we determine the X-ray structures of PilT in four unique conformations and use these structures to classify the conformation of available PilT/VirB11-like family member structures. Single particle electron cryomicroscopy (cryoEM) structures of PilT reveal condition-dependent preferences for C<sub>2,</sub> C<sub>3</sub>, and C<sub>6</sub> conformations. The physiologic importance of these conformations is validated by coevolution analysis and functional studies of point mutants, identifying a rare gain-of-function mutation that favours the C<sub>2</sub> conformation. With these data, we propose a comprehensive model of PilT function with broad implications for PilT/VirB11-like family members.

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