Biogenesis of E. coli Pap pili: PapH, a minor pilin subunit involved in cell anchoring and length modulation

Båga, Monica; Norgren, Mari; Normark, Staffan

doi:10.1016/0092-8674(87)90565-4

Cited by 160 publications

(125 citation statements)

References 42 publications

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“…These results further suggest that the pore gating function of the PLUG domain can be decoupled from its function in catalysis. Previous studies revealed the importance of PapH (the terminator subunit of P pili) in anchoring and terminating P pili at the OM (18,28). Biophysical studies further revealed that the PapDH complex was specifically recruited by the PLUG domain or the intramolecular NTD/PLUG complex but did not bind to CTD2 (23).…”

Section: Resultsmentioning

confidence: 98%

“…The P pilus comprises a flexible-tip fibrillum made up of minor pilins with the two-domain adhesin PapG at the distal end where it can recognize Galα1-4Gal disaccharide-containing glycolipids found in the human kidney (14,15). The P pilus tip is joined to a righthanded, helical pilus rod made up of PapA pilins (16,17) and is anchored in the OM via the terminator/anchoring subunit PapH (18). Type 1 pili are composed of FimA subunits making up the pilus rod joined to the tip fibrillum, which contains the FimH tip adhesin joined to FimG and adapted to the FimA rod by FimF.…”

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confidence: 99%

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Molecular basis of usher pore gating in Escherichia coli pilus biogenesis

Volkan

Kalas

Pinkner³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Extracellular fibers called chaperone-usher pathway pili are critical virulence factors in a wide range of Gram-negative pathogenic bacteria that facilitate binding and invasion into host tissues and mediate biofilm formation. Chaperone-usher pathway ushers, which catalyze pilus assembly, contain five functional domains: a 24-stranded transmembrane β-barrel translocation domain (TD), a β-sandwich plug domain (PLUG), an N-terminal periplasmic domain, and two Cterminal periplasmic domains (CTD1 and 2). Pore gating occurs by a mechanism whereby the PLUG resides stably within the TD pore when the usher is inactive and then upon activation is translocated into the periplasmic space, where it functions in pilus assembly. Using antibiotic sensitivity and electrophysiology experiments, a single salt bridge was shown to function in maintaining the PLUG in the TD channel of the P pilus usher PapC, and a loop between the 12th and 13th beta strands of the TD (β12-13 loop) was found to facilitate pore opening. Mutation of the β12-13 loop resulted in a closed PapC pore, which was unable to efficiently mediate pilus assembly. Deletion of the PapH terminator/anchor resulted in increased OM permeability, suggesting a role for the proper anchoring of pili in retaining OM integrity. Further, we introduced cysteine residues in the PLUG and N-terminal periplasmic domains that resulted in a FimD usher with a greater propensity to exist in an open conformation, resulting in increased OM permeability but no loss in type 1 pilus assembly. These studies provide insights into the molecular basis of usher pore gating and its roles in pilus biogenesis and OM permeability.outer membrane usher | macromolecular assembly | bacterial pathogenesis

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Section: Resultsmentioning

confidence: 98%

mentioning

confidence: 99%

Molecular basis of usher pore gating in Escherichia coli pilus biogenesis

Volkan

Kalas

Pinkner³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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“…Pap (type I) pili are assembled from six pilins, all of which are encoded by a single large operon (Soto and Hultgren 1999). To identify the position of pilins within the pilus, wild-type E. coli and mutants lacking single pilin genes were analyzed by electron microscopy and immuno-gold labeling (Norgren et al 1984;Baga et al 1987;Lindberg et al 1987;Kuehn et al 1992;Jacob-Dubuisson et al 1993). PapG pilin (G in Fig.…”

Section: Assembly Of Type I Pilimentioning

confidence: 99%

Protein secretion and the pathogenesis of bacterial infections

Lee¹,

Schneewind²

2001

Genes Dev.

202

155

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“…Upon their transfer to the periplasm (10), these structural subunits are assembled through the actions of the periplasmic chaperone (PapD) and the integral outer membrane usher (PapC) in a specific order into bipartite P pilus fibers composing an open helical tip fibrillum made up of (from the tip) PapG (11), PapF, multiple PapEs, and PapK (12,13), connected to a right-handed helical rod made up of thousands of copies of PapA (14)(15)(16)(17) and terminated by a single copy of PapH (18,19). Each pilin Ig fold is missing the seventh C-terminal β-strand of the canonical Ig fold.…”

mentioning

confidence: 99%

Domain activities of PapC usher reveal the mechanism of action of an Escherichia coli molecular machine

Volkan¹,

Ford

Pinkner³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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P pili are prototypical chaperone-usher pathway-assembled pili used by Gram-negative bacteria to adhere to host tissues. The PapC usher contains five functional domains: a transmembrane β-barrel, a β-sandwich Plug, an N-terminal (periplasmic) domain (NTD), and two C-terminal (periplasmic) domains, CTD1 and CTD2. Here, we delineated usher domain interactions between themselves and with chaperone-subunit complexes and showed that overexpression of individual usher domains inhibits pilus assembly. Prior work revealed that the Plug domain occludes the pore of the transmembrane domain of a solitary usher, but the chaperone-adhesinbound usher has its Plug displaced from the pore, adjacent to the NTD. We demonstrate an interaction between the NTD and Plug domains that suggests a biophysical basis for usher gating. Furthermore, we found that the NTD exhibits high-affinity binding to the chaperone-adhesin (PapDG) complex and low-affinity binding to the major tip subunit PapE (PapDE). We also demonstrate that CTD2 binds with lower affinity to all tested chaperone-subunit complexes except for the chaperone-terminator subunit (PapDH) and has a catalytic role in dissociating the NTD-PapDG complex, suggesting an interplay between recruitment to the NTD and transfer to CTD2 during pilus initiation. The Plug domain and the NTDPlug complex bound all of the chaperone-subunit complexes tested including PapDH, suggesting that the Plug actively recruits chaperone-subunit complexes to the usher and is the sole recruiter of PapDH. Overall, our studies reveal the cooperative, active roles played by periplasmic domains of the usher to initiate, grow, and terminate a prototypical chaperone-usher pathway pilus.biolayer interferometry | macromolecular assembly | urinary tract infections | virulence factor | bacterial pathogenesis

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Biogenesis of E. coli Pap pili: PapH, a minor pilin subunit involved in cell anchoring and length modulation

Cited by 160 publications

References 42 publications

Molecular basis of usher pore gating in Escherichia coli pilus biogenesis

Molecular basis of usher pore gating in Escherichia coli pilus biogenesis

Protein secretion and the pathogenesis of bacterial infections

Domain activities of PapC usher reveal the mechanism of action of an Escherichia coli molecular machine

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