Functional role of the type 1 pilus rod structure in mediating host-pathogen interactions

Spaulding, Caitlin N.; Schreiber, Henry L.; Zheng, Weili; Dodson, Karen W.; Hazen, Jennie E; Conover, Matt S.; Wang, Fengbin; Svenmarker, Pontus; Luna-Rico, Areli; Francetić, Olivera; Andersson, Magnus; Hultgren, Scott J.

doi:10.7554/elife.31662

Cited by 79 publications

(65 citation statements)

References 81 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Similar local strand unwinding has been observed for chaperone-usher pili. Near-atomic resolution structural analysis of the corresponding pili with cryo-EM revealed that the unwinding of the helical assembly occurred analogously at the weaker axial interface (44)(45)(46).…”

Section: Discussionmentioning

confidence: 99%

Ambidextrous helical nanotubes from self-assembly of designed helical hairpin motifs

Hughes

Wang

et al. 2019

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

Self Cite

View full text Add to dashboard Cite

Tandem repeat proteins exhibit native designability and represent potentially useful scaffolds for the construction of synthetic biomimetic assemblies. We have designed 2 synthetic peptides, HEAT_R1 and LRV_M3Δ1, based on the consensus sequences of single repeats of thermophilic HEAT (PBS_HEAT) and Leucine-Rich Variant (LRV) structural motifs, respectively. Self-assembly of the peptides afforded high-aspect ratio helical nanotubes. Cryo-electron microscopy with direct electron detection was employed to analyze the structures of the solvated filaments. The 3D reconstructions from the cryo-EM maps led to atomic models for the HEAT_R1 and LRV_M3Δ1 filaments at resolutions of 6.0 and 4.4 Å, respectively. Surprisingly, despite sequence similarity at the lateral packing interface, HEAT_R1 and LRV_M3Δ1 filaments adopt the opposite helical hand and differ significantly in helical geometry, while retaining a local conformation similar to previously characterized repeat proteins of the same class. The differences in the 2 filaments could be rationalized on the basis of differences in cohesive interactions at the lateral and axial interfaces. These structural data reinforce previous observations regarding the structural plasticity of helical protein assemblies and the need for high-resolution structural analysis. Despite these observations, the native designability of tandem repeat proteins offers the opportunity to engineer novel helical nanotubes. Moreover, the resultant nanotubes have independently addressable and chemically distinguishable interior and exterior surfaces that would facilitate applications in selective recognition, transport, and release.

show abstract

Section: Discussionmentioning

confidence: 99%

Ambidextrous helical nanotubes from self-assembly of designed helical hairpin motifs

Hughes

Wang

et al. 2019

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

Self Cite

View full text Add to dashboard Cite

show abstract

“…Interestingly, such experiments showed that type 1 pilus rods require slightly higher forces to induce rod unwinding than do P pilus rods, prompting the suggestion that type 1 pili are better adapted to withstand the more turbulent flows of the lower urinary tract (36,76). Several structures of the type 1 pilus rod have been determined (10,77,78) (Fig. 3), revealing a very similar overall architecture to the P pilus rod, except that the type 1 pilus rod lacks the so-called "staple" region (9) (Fig.…”

Section: Clinging On: the Importance Of The Pilus Rodmentioning

confidence: 99%

The Remarkable Biomechanical Properties of the Type 1 Chaperone-Usher Pilus: A Structural and Molecular Perspective

Hospenthal

Waksman

2019

Microbiol Spectr

View full text Add to dashboard Cite

Chaperone-usher (CU) pili are long, supramolecular protein fibers tethered to the surface of numerous bacterial pathogens. These virulence factors function primarily in bacterial adhesion to host tissues, but they also mediate biofilm formation. Type 1 and P pili of uropathogenic Escherichia coli (UPEC) are the two best-studied CU pilus examples, and here we primarily focus on the former. UPEC can be transmitted to the urinary tract by fecal shedding. It can then ascend up the urinary tract and cause disease by invading and colonizing host tissues of the bladder, causing cystitis, and the kidneys, causing pyelonephritis. FimH is the subunit displayed at the tip of type 1 pili and mediates adhesion to mannosylated host cells via a unique catch-bond mechanism. In response to shear forces caused by urine flow, FimH can transition from a low-affinity to high-affinity binding mode. This clever allosteric mechanism allows UPEC cells to remain tightly attached during periods of urine flow, while loosening their grip to allow dissemination through the urinary tract during urine stasis. Moreover, the bulk of a CU pilus is made up of the rod, which can reversibly uncoil in response to urine flow to evenly spread the tensile forces over the entire pilus length. We here explore the novel structural and mechanistic findings relating to the type 1 pilus FimH catch-bond and rod uncoiling and explain how they function together to enable successful attachment, spread, and persistence in the hostile urinary tract.

show abstract

“…Crude sheared fractions of strains BW25113 F′ expressing ppdD (from pCHAP8565) and either the T2SS genes (from pCHAP8184) (panels A‐C) or T4PS genes (from pMS41) (panels D‐F) were analysed. Black arrows indicate Type 1 pili (in panels B and C) as shown previously in (Spaulding et al, ), the blue arrow indicates flagella (in panel E) and white arrows indicate PpdD T4P. The scale bar (A) is 50 nm.…”

Section: Resultsmentioning

confidence: 99%

Functional reconstitution of the type IVa pilus assembly system from enterohaemorrhagic Escherichia coli

Rico

Zheng

Petiot

et al. 2019

Molecular Microbiology

Self Cite

View full text Add to dashboard Cite

Summary Type 4a pili (T4aP) are long, thin and dynamic fibres displayed on the surface of diverse bacteria promoting adherence, motility and transport functions. Genomes of many Enterobacteriaceae contain conserved gene clusters encoding putative T4aP assembly systems. However, their expression has been observed only in few strains including Enterohaemorrhagic Escherichia coli (EHEC) and their inducers remain unknown. Here we used EHEC genomic DNA as a template to amplify and assemble an artificial operon composed of four gene clusters encoding 13 pilus assembly proteins. Controlled expressions of this operon in nonpathogenic E. coli strains led to efficient assembly of T4aP composed of the major pilin PpdD, as shown by shearing assays and immunofluorescence microscopy. When compared with PpdD pili assembled in a heterologous Klebsiella T2SS type 2 secretion system (T2SS) by using cryo‐electron microscopy (cryoEM), these pili showed indistinguishable helical parameters, emphasizing that major pilins are the principal determinants of the fibre structure. Bacterial two‐hybrid analysis identified several interactions of PpdD with T4aP assembly proteins, and with components of the T2SS that allow for heterologous fibre assembly. These studies lay ground for further characterization of the T4aP structure, function and biogenesis in enterobacteria.

show abstract

Functional role of the type 1 pilus rod structure in mediating host-pathogen interactions

Cited by 79 publications

References 81 publications

Ambidextrous helical nanotubes from self-assembly of designed helical hairpin motifs

Ambidextrous helical nanotubes from self-assembly of designed helical hairpin motifs

The Remarkable Biomechanical Properties of the Type 1 Chaperone-Usher Pilus: A Structural and Molecular Perspective

Functional reconstitution of the type IVa pilus assembly system from enterohaemorrhagic Escherichia coli

Contact Info

Product

Resources

About