Pseudomonas aeruginosa Minor Pilins Prime Type IVa Pilus Assembly and Promote Surface Display of the PilY1 Adhesin

Nguyen, Ylan; Sugiman-Marangos, Seiji; Harvey, Hanjeong; Bell, S.; Charlton, Carmen; Junop, M.S.; Burrows, Lori L.

doi:10.1074/jbc.m114.616904

Cited by 120 publications

(159 citation statements)

References 67 publications

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“…2a). The OM secretin subcomplex includes PilQ, a multimeric OM pore protein through which the growing pilus is assembled and disassembled [65][66][67][68] , and the pilotin protein The final structural component completing the T4P system is the helical pilus filament, composed of major and minor pilin subunits and adhesin molecules 81,82 (FIG. 2d).…”

Section: T4p Biogenesismentioning

confidence: 99%

A comprehensive guide to pilus biogenesis in Gram-negative bacteria

2017

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Pili are critical virulence factors of many Gram-negative pathogens. These surface structures provide bacteria with a link to their outside environments, allowing bacteria to interact with their hosts, other surfaces or with each other. They can even provide a conduit for the exchange of information. The surface chemistries and other biophysical properties of pili are uniquely adapted to the environmental challenges faced by bacteria. The assembly of these surface structures presents a molecular engineering challenge, which bacteria have solved in a variety of unique ways. In this review, we examine recent advances in our structural understanding of various Gram-negative pilus systems and discuss their functional implications.

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Section: T4p Biogenesismentioning

confidence: 99%

A comprehensive guide to pilus biogenesis in Gram-negative bacteria

2017

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“…There are two main classes of T4P, type IVa and type IVb, distinguished by differences in their subunits and assembly machineries (1,3,4). The pili are composed of thousands of major pilin subunits, but minor (low abundance) pilin subunits are also present, potentially at the tip of the pilus due to their role in priming of pilus assembly (5)(6)(7)(8)(9)(10).…”

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confidence: 99%

Structural and Functional Studies of the Pseudomonas aeruginosa Minor Pilin, PilE

Nguyen

Harvey

Sugiman-Marangos

et al. 2015

Journal of Biological Chemistry

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“…The secretin and motor subcomplexes are linked by the alignment subcomplex (PilMNOP), which may also have a role in secretin gating (24,25). The final subcomplex is the pilus, a helical filament of major and minor pilin subunits plus the PilY1 adhesin (13,18,(26)(27)(28). Together with a number of regulatory proteins whose functions are incompletely understood (29)(30)(31)(32), these subcomplexes form a fully functional T4P system.…”

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confidence: 99%

Novel Role for PilNO in Type IV Pilus Retraction Revealed by Alignment Subcomplex Mutations

et al. 2015

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Type IV pili (T4P) are dynamic protein filaments that mediate bacterial adhesion, biofilm formation, and twitching motility. The highly conserved PilMNOP proteins form an inner membrane alignment subcomplex required for function of the T4P system, though their exact roles are unclear. Three potential interaction interfaces for PilNO were identified: core-core, coiled coils (CC), and the transmembrane segments (TMSs). A high-confidence PilNO heterodimer model was used to select key residues for mutation, and the resulting effects on protein-protein interactions were examined both in a bacterial two-hybrid (BTH) system and in their native Pseudomonas aeruginosa context. Mutations in the oppositely charged CC regions or the TMS disrupted PilNO heterodimer formation in the BTH assay, while up to six combined mutations in the core failed to disrupt the interaction. When the mutations were introduced into the P. aeruginosa chromosome at the pilN or pilO locus, specific changes at each of the three interfaces-including core mutations that failed to disrupt interactions in the BTH system-abrogated surface piliation and/or impaired twitching motility. Unexpectedly, specific CC mutants were hyperpiliated but nonmotile, a hallmark of pilus retraction defects. These data suggest that PilNO participate in both the extension and retraction of T4P. Our findings support a model of multiple, precise interaction interfaces between PilNO; emphasize the importance of studying protein function in a minimally perturbed context and stoichiometry; and highlight potential target sites for development of small-molecule inhibitors of the T4P system. IMPORTANCEPseudomonas aeruginosa is an opportunistic pathogen that uses type IV pili (T4P) for host attachment. The T4P machinery is composed of four cell envelope-spanning subcomplexes. PilN and PilO heterodimers are part of the alignment subcomplex and essential for T4P function. Three potential PilNO interaction interfaces (the core-core, coiled-coil, and transmembrane segment interfaces) were probed using site-directed mutagenesis followed by functional assays in an Escherichia coli two-hybrid system and in P. aeruginosa. Several mutations blocked T4P assembly and/or motility, including two that revealed a novel role for PilNO in pilus retraction, while other mutations affected extension dynamics. These critical PilNO interaction interfaces represent novel targets for small-molecule inhibitors with the potential to disrupt T4P function.A larming increases in antibiotic resistance have prompted the search for alternative strategies to combat bacterial infection (1). Genomic strategies designed to identify essential bacterial targets have been informative but less successful for drug development than originally predicted due to high mutation rates and resistance (2). Alternative strategies-such as phage therapy and the use of "antivirulence" drugs that target bacterial factors involved in pathogenesis-show promise since they can effectively disarm bacteria, which are then more easily...

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Pseudomonas aeruginosa Minor Pilins Prime Type IVa Pilus Assembly and Promote Surface Display of the PilY1 Adhesin

Cited by 120 publications

References 67 publications

A comprehensive guide to pilus biogenesis in Gram-negative bacteria

A comprehensive guide to pilus biogenesis in Gram-negative bacteria

Structural and Functional Studies of the Pseudomonas aeruginosa Minor Pilin, PilE

Novel Role for PilNO in Type IV Pilus Retraction Revealed by Alignment Subcomplex Mutations

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